Method of producing pyrone and pyridone derivatives

ABSTRACT

The present invention provides a pyrone derivative and a pyridone derivative, which are novel intermediates for synthesizing an anti-influenza drug, a method of producing the same, and a method of using the same.

TECHNICAL FIELD

The present invention relates to a pyrone derivative and a pyridonederivative, which are novel intermediates for synthesizing ananti-influenza drug exhibiting the cap-dependent endonuclease inhibitoryactivity, a method of producing the same, and a method of using thesame.

BACKGROUND ART

Influenza is an acute respiratory infectious disease caused by infectionwith an influenza virus. In Japan, there is a report of a few millionsof influenza-like patients every winter, and influenza is accompaniedwith high morbidity and mortality. Influenza is a particularly importantdisease in a high risk population such as baby and elderly, acomplication rate with pneumonia is high in elderly, and death withinfluenza is occupied with elderly in many cases.

As anti-influenza drugs, Symmetrel (trade name: Amantadine) andFlumadine (trade name: Rimantadine) which inhibit the denucleationprocess of a virus, and Oseltamivir (trade name: Tamiflu) and Zanamivir(trade name: Relenza) which are neuraminidase inhibitors suppressingvirus budding and release from a cell are known. However, since problemsof appearances of resistant strains and side effects, and worldwideepidemic of a new-type influenza virus having high pathogenicity andmortality are feared, development of an anti-influenza drug having anovel mechanism has been desired.

Since a cap-dependent endonuclease which is an influenza virus-derivedenzyme is essential for virus proliferation, and has the virus-specificenzymatic activity which is not possessed by a host, it is believed thatthe endonuclease is suitable for a target of an anti-influenza drug. Thecap-dependent endonuclease has a host mRNA precursor as a substrate, andhas the endonuclease activity of producing a fragment of 9 to 13 basesincluding a cap structure (not including the number of bases of the capstructure). This fragment functions as a primer of a virus RNApolymerase, and is used in synthesizing mRNA encoding a virus protein.That is, it is believed that a substance which inhibits thecap-dependent endonuclease inhibits synthesis of a virus protein byinhibiting synthesis of virus mRNA and, as a result, inhibits virusproliferation.

As the substance which inhibits the cap-dependent endonuclease,flutamide (Patent Document 1 and Non-Patent Documents 1 and 2) and4-substituted 2,4-dioxobutanoic acid (Non-Patent Documents 3 to 5) arereported, but they have not yet led to clinical use as anti-influenzadrugs. In addition, Patent Documents 2 to 11 and Non-Patent Document 6describe compounds having a similar structure to that of a novelanti-influenza drug exhibiting the cap-dependent endonuclease inhibitoryactivity, and a method of producing the same. In addition, a pyronederivative and a pyridone derivative are disclosed in Non-PatentDocuments 7 to 9.

PRIOR ART DOCUMENTS Patent Documents

-   [Patent Document 1] GB No. 2280435 specification-   [Patent Document 2] International Publication No. 2007/049675    pamphlet-   [Patent Document 3] International Publication No. 2006/088173    pamphlet-   [Patent Document 4] International Publication No. 2006/066414    pamphlet-   [Patent Document 5] International Publication No. 2005/092099    pamphlet-   [Patent Document 6] International Publication No. 2005/087766    pamphlet-   [Patent Document 7] International Publication No. 2005/016927    pamphlet-   [Patent Document 8] International Publication No. 2004/024078    pamphlet-   [Patent Document 9] International Publication No. 2006/116764    pamphlet-   [Patent Document 10] International Publication No. 2006/030807    pamphlet-   [Patent Document 11] Japanese Patent Laid-open Publication No.    2006-342115

Non-Patent Documents

-   [Non-Patent Document 1] Tetrahedron Lett 1995, 36(12), 2005-   [Non-Patent Document 2] Tetrahedron Lett 1995, 36(12), 2009-   [Non-Patent Document 3] Antimicrobial Agents And Chemotherapy,    December 1994, p. 2827-2837-   [Non-Patent Document 4] Antimicrobial Agents And Chemotherapy, May    1996, p. 1304-1307-   [Non-Patent Document 5] J. Med. Chem. 2003, 46, 1153-1164-   [Non-Patent Document 6] Bioorganic & Medicinal Chemistry Letters    17 (2007) 5595-5599-   [Non-Patent Document 7] Journal of Organic Chemistry 1960, 25, p.    1052-1053-   [Non-Patent Document 8] Tetrahedron Lett 1981, 22(23), 2207-   [Non-Patent Document 9] Journal of Organic Chemistry 1991, 56, p.    4963-4967

SUMMARY OF THE INVENTION Problems to be Solved by the Invention

An object of the present invention is to provide a novel pyronederivative and a novel pyridone derivative, which are novelintermediates for synthesizing an anti-influenza drug, a method ofproducing the same, and a method of using the same. Specifically, theobject is to efficiently produce compounds and the like useful as ananti-influenza drug, which are exemplified by formula (I), formula (II)or formula (III):

(wherein R¹ is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkylamino optionally substituted by substituent E, orheterocyclyl lower alkylamino optionally substituted by substituent E,

Z is CR¹²R¹³, or a single bond,

Y is CH₂, an oxygen atom, or N—R¹⁴,

R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are eachindependently hydrogen, lower alkyl optionally substituted bysubstituent E, lower alkenyl optionally substituted by substituent E,lower alkynyl optionally substituted by substituent E, carbocyclyl loweralkyl optionally substituted by substituent E, or heterocyclyl loweralkyl optionally substituted by substituent E,

R³ and R⁵ may be taken together to form 4 to 8-membered carbocyclyloptionally substituted by substituent E, or 4 to 8-membered heterocyclyloptionally substituted by substituent E, or R⁷ and R⁵ may be takentogether to form 4 to 8-membered heterocyclyl optionally substituted bysubstituent E,

wherein when Z is a single bond, R¹⁴ and R¹⁰ may be taken together toform 4 to 8-membered heterocyclyl optionally substituted by substituentE. Substituent E; halogen, cyano, hydroxy, carboxy, formyl, amino, oxo,nitro, lower alkyl, halogeno lower alkyl, lower alkyloxy, carbocyclyloptionally substituted by substituent F, heterocyclyl optionallysubstituted by substituent F, carbocyclyl lower alkyloxy optionallysubstituted by substituent F, heterocyclyl lower alkyloxy optionallysubstituted by substituent F, carbocyclyl lower alkylthio optionallysubstituted by substituent F, heterocyclyl lower alkylthio optionallysubstituted by substituent F, carbocyclyl lower alkylamino optionallysubstituted by substituent F, heterocyclyl lower alkylamino optionallysubstituted by substituent F, carbocyclyloxy optionally substituted bysubstituent F, heterocyclyloxy optionally substituted by substituent F,carbocyclylcarbonyl optionally substituted by substituent F,heterocyclylcarbonyl optionally substituted by substituent F,carbocyclylaminocarbonyl optionally substituted by substituent F,heterocyclylaminocarbonyl optionally substituted by substituent F,halogeno lower alkyloxy, lower alkyloxy lower alkyl, lower alkyloxylower alkyloxy, lower alkylcarbonyl, lower alkyloxycarbonyl, loweralkyloxycarbonylamino, lower alkylamino, lower alkylcarbonylamino, loweralkylaminocarbonyl, lower alkylsulfonyl, and lower alkylsulfonylamino;

Substituent F: halogen, hydroxy, carboxy, amino, oxo, nitro, loweralkyl, halogeno lower alkyl, lower alkyloxy, and amino protectivegroup).

Means for Solving the Problems

The present invention provides the following items.

(Item 1)

A method of producing a compound shown by formula (X4) or a saltthereof, comprising steps of

(Step B)

reacting a compound shown by formula (X2):

(wherein R^(1d) is hydrogen, halogen, lower alkyloxy optionallysubstituted by substituent E, carbocyclyl lower alkyloxy optionallysubstituted by substituent E, heterocyclyl lower alkyloxy optionallysubstituted by substituent E, or —OSi(R^(1e))₃,

R^(1e)s are each independently lower alkyl optionally substituted bysubstituent E, carbocyclyl optionally substituted by substituent E,heterocyclyl optionally substituted by substituent E, carbocyclyl loweralkyl optionally substituted by substituent E or heterocyclyl loweralkyl optionally substituted by substituent E,

R^(2d) is hydrogen, lower alkyl optionally substituted by substituent E,carbocyclyl lower alkyl optionally substituted by substituent E, orheterocyclyl lower alkyl optionally substituted by substituent E,

R^(3d) is hydrogen, lower alkyl optionally substituted by substituent E,—N(R^(3e))₂, or —OR^(3e),

R^(3e)s are each independently lower alkyl optionally substituted bysubstituent E, or may be taken together to form a heterocycle, and

wavy line is E form and/or Z form or the mixture thereof.

Substituent E: halogen, cyano, hydroxy, carboxy, formyl, amino, oxo,nitro, lower alkyl, halogeno lower alkyl, lower alkyloxy, carbocyclyloptionally substituted by substituent F, heterocyclyl optionallysubstituted by substituent F, carbocyclyl lower alkyloxy optionallysubstituted by substituent F, heterocyclyl lower alkyloxy optionallysubstituted by substituent F, carbocyclyl lower alkylthio optionallysubstituted by substituent F, heterocyclyl lower alkylthio optionallysubstituted by substituent F, carbocyclyl lower alkylamino optionallysubstituted by substituent F, heterocyclyl lower alkylamino optionallysubstituted by substituent F, carbocyclyloxy optionally substituted bysubstituent F, heterocyclyloxy optionally substituted by substituent F,carbocyclylcarbonyl optionally substituted by substituent F,heterocyclylcarbonyl optionally substituted by substituent F,carbocyclylaminocarbonyl optionally substituted by substituent F,heterocyclylaminocarbonyl optionally substituted by substituent F,halogeno lower alkyloxy, lower alkyloxy lower alkyl, lower alkyloxylower alkyloxy, lower alkylcarbonyl, lower alkyloxycarbonyl, loweralkyloxycarbonylamino, lower alkylamino, lower alkylcarbonylamino, loweralkylaminocarbonyl, lower alkylsulfonyl, and lower alkylsulfonylamino;Substituent F: halogen, hydroxy, carboxy, amino, oxo, nitro, loweralkyl, halogeno lower alkyl, lower alkyloxy, and amino protective group)with a compound shown by formula (V2):

(wherein R^(4d) is lower alkyl optionally substituted by substituent E,carbocyclyl lower alkyl optionally substituted by substituent E, orheterocyclyl lower alkyl optionally substituted by substituent E,

R^(5d) is hydrogen, halogen, lower alkyloxy optionally substituted bysubstituent E, or —O—SO₂—R^(5e),

R^(5e) is lower alkyl optionally substituted by substituent E,carbocyclyl optionally substituted by substituent E, heterocyclyloptionally substituted by substituent E, carbocyclyl lower alkyloptionally substituted by substituent E, or heterocyclyl lower alkyloptionally substituted by substituent E, and

substituent E is defined above)

to obtain a compound shown by formula (X3):

(wherein each symbol is defined above); and

(Step C)

reacting the compound shown by formula (X3) with a compound shown byformula (V3):

[Chemical formula 5]

H₂N—R^(6d)  (V3)

(wherein R^(6d) is lower alkyl optionally substituted by substituent E,lower alkenyl optionally substituted by substituent E, amino optionallysubstituted by substituent E, lower alkylamino optionally substituted bysubstituent E, carbocyclyl optionally substituted by substituent E, orheterocyclyl optionally substituted by substituent E, and

substituent E is defined above)

to obtain a compound shown by formula (X4):

(wherein each symbol is defined above).

(Item 2)

A method according to Item 1, wherein Step B and Step C are continuouslyperformed.

(Item 3)

A method of producing a compound shown by formula (XA4), or a saltthereof:

(wherein R^(a) is hydrogen, hydroxy, lower alkylamino optionallysubstituted by substituent E, lower alkenylamino optionally substitutedby substituent E, lower alkynylamino optionally substituted bysubstituent E, carbocyclyl lower alkylamino optionally substituted bysubstituent E, or heterocyclyl lower alkylamino optionally substitutedby substituent E, and

each symbol is defined above)

comprising the step of:

reacting a compound shown by formula (X2):

(wherein each symbol is defined above)with a compound shown by formula (VA2):

(wherein R^(4e)s are each independently hydrogen, lower alkyl optionallysubstituted by substituent E, carbocyclyl optionally substituted bysubstituent E, or heterocyclyl optionally substituted by substituent E,and

R^(5d) and substituent E are defined above); and

reacting with a compound shown by formula (V3):

[Chemical formula 9]

H₂N—R^(6d)  (V3)

(wherein R^(6d) is defined above).

(Item 4)

A method of producing a compound shown by formula (X3) or formula (XA3),or a salt thereof:

(wherein each symbol is defined in Item 1)comprising reacting a compound shown by formula (X2):

(wherein each symbol is defined in Item 1)with a compound shown by formula (V2) or formula (VA2):

(wherein each symbol is defined in Item 1).

(Item 5)

A method of producing a compound shown by formula (X4) or formula (XA4),or a salt thereof:

(wherein each symbol is defined in Item 1 and Item 3)comprising the step of:

reacting a compound shown by formula (X3) or formula (XA3):

(wherein each symbol is defined in Item 1)or the derivative of formula (XA3) with a compound shown by formula(V3):

[Chemical formula 15]

H₂N—R^(6d)  (V3)

(wherein each symbol is defined in Item 1).

(Item 6)

A method of producing a compound shown by formula (X4′) or formula(XA4′), or a salt thereof:

(wherein each symbol is defined in Item 1)comprising reacting a compound shown by formula (X2):

(wherein each symbol is defined in Item 1)with a compound shown by formula (V2) or formula (VA2):

(wherein each symbol is defined in Item 1)and a compound shown by formula (V2′):

[Chemical formula 19]

NH⁴⁺X^(d−)  (V2′)

(wherein X^(d−) is counter anion of ammonium cation).

(Item 7)

A method of producing a compound shown by formula (X4) or formula (XA4),or a salt thereof:

(wherein each symbol is defined in Item 1 or Item 3)comprising the step of:

reacting the compound shown by formula (X4′) or formula (XA4′):

(wherein each symbol is defined in Item 1)obtained in the production method as defined in Item 6, or thederivative of formula (XA4′) with a compound shown by formula (V3′):

[Chemical formula 22]

R^(6d)-L^(d)  (V3′)

(wherein R^(6d) is defined in Item 1,

L^(d) is a leaving group, and Ph is a phenyl group).

(Item 8)

A method according to Item 1, 2, 3, 4 or Item 6, wherein the compoundshown by formula (X2) is obtained by reacting a compound shown byformula (X1):

(wherein each symbol is defined in Item 1)with a compound shown by formula (V1):

(wherein P^(d) is lower alkyl optionally substituted by substituent E,and R^(3d) and substituent E are defined in Item 1).

(Item 9)

A method according to Item 1, 2, 3, 4 or Item 6, wherein the compoundshown by formula (X2) is obtained by reacting a compound shown byformula (Z1):

(wherein each symbol is defined in Item 1)with a compound shown by formula (Z2):

(wherein each symbol is defined in Item 1).

(Item 10)

A compound shown by formula (X3), or a pharmaceutically acceptable saltthereof or solvate thereof:

(wherein each symbol is defined in Item 1).

(Item 11)

A compound shown by formula (X4), or a pharmaceutically acceptable saltthereof or solvate thereof:

(wherein each symbol is defined in Item 1).

(Item 12)

A compound shown by formula (X4′), or a pharmaceutically acceptable saltthereof or solvate thereof:

(wherein each symbol is defined in claim 1).

(Item 13)

A crystal of a compound shown by formula (1D) or a solvate thereof:

(wherein Me is a methyl group, and Bn is a benzyl group),wherein the compound has a peak at a diffraction angle (2θ): 7.9°±0.2°,10.0°±0.2°, 11.5°±0.2°, 20.0°±0.2°, 23.4°±0.2° and 34.0°±0.2° in apowder X-ray diffraction spectrum.

(Item 14)

A crystal of a compound shown by formula (2D) or a solvate thereof:

(wherein Me is a methyl group, Et is an ethyl group, and Bn is a benzylgroup),wherein the compound has a peak at a diffraction angle (2θ): 17.6°±0.2°,25.2°±0.2°, 26.4°±0.2° and 28.1°±0.2° in a powder X-ray diffractionspectrum.

(Item 15)

A crystal of a compound shown by formula (9C′) or a solvate thereof:

(wherein Me is a methyl group, and Bn is a benzyl group),wherein the compound has a peak at a diffraction angle (2θ): 14.2°±0.2°,16.0°±0.2°, 22.0°±0.2°, 22.2°±0.2°, 24.4°±0.2° and 25.9°±0.2° in apowder X-ray diffraction spectrum.

(Item 16)

A crystal according to Item 13, which is characterized by a powder X-raydiffraction spectrum which is substantially consistent with FIG. 1.

(Item 17)

A crystal according to Item 14, which is characterized by a powder X-raydiffraction spectrum which is substantially consistent with FIG. 2.

(Item 18)

A crystal according to Item 15, which is characterized by a powder X-raydiffraction spectrum which is substantially consistent with FIG. 3.

Effect of the Invention

The production method according to the present invention is a methodthat can produce compounds included in the formula (I), the formula (II)or the formula (III) etc., which are novel anti-influenza drugs, at ahigh yield, efficiently, and/or in a short step. In addition, byperforming the production method according to the present invention,there are a plurality of advantages that use of a reaction reagentaccompanying with toxicity can be avoided, a reaction accompanying witha risk can be avoided, use of an expensive reaction reagent can beavoided, and use of an environmentally harmful reagent and solvent canbe avoided, etc. The pyrone derivative (X3), and the pyridone derivative(X4) and/or (X4′) which are the present compound have versatility, andare useful as common intermediates at production of the compoundsincluded in the formula (I), the formula (II) or the formula (III).Furthermore, the pyrone derivative (X3) and/or the pyridone derivative(X4) can be also obtained as a crystal. Since these crystals have smallhygroscopy, and exhibit high stability to light and heat, they haveadvantages that storage and handling in production are excellent and thelike. Therefore, the production method as well as intermediates andcrystals thereof according to the present invention are useful inindustrially producing medicaments (anti-influenza drugs, anti-HIVdrugs, anti-inflammatory drugs, tranquilizers, anti-tumor drugs etc.).

(wherein each symbol is defined above)

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a powder X-ray pattern of Compound 1D obtained in Example 1.An ordinate indicates a peak intensity, and an abscissa indicates adiffraction angle (2θ).

FIG. 2 is a powder X-ray pattern of Compound 2D obtained in Example 2.An ordinate indicates a peak intensity, and an abscissa indicates adiffraction angle (2θ).

FIG. 3 is a powder X-ray pattern of Compound 9C′ obtained in Example 9.An ordinate indicates a peak intensity, and an abscissa indicates adiffraction angle (2θ).

MODE FOR CARRYING OUT THE INVENTION

In the present specification, the “halogen” contains a fluorine atom, achlorine atom, a bromine atom and an iodine atom.

In the present specification, the “lower alkyl” includes straight orbranched alkyl having a carbon number of 1 to 15, preferably a carbonnumber of 1 to 10, more preferably a carbon number of 1 to 6, furtherpreferably a carbon number of 1 to 4, and examples thereof includemethyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, isohexyl, n-heptyl,isoheptyl, n-octyl, isooctyl, n-nonyl and n-decyl.

Examples of a preferable aspect of the “lower alkyl” include methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl andn-pentyl. Examples of a further preferable aspect include methyl, ethyl,n-propyl, isopropyl and tert-butyl.

In the present specification, the “lower alkenyl” includes straight orbranched alkenyl having one or more double bonds at arbitrary positionsand having 2 to 15 carbon atoms, preferably 2 to 10 carbon atoms, morepreferably 2 to 6 carbon atoms, further preferably 2 to 4 carbon atoms.Specifically, examples thereof include vinyl, allyl, propenyl,isopropenyl, butenyl, isobutenyl, prenyl, butadienyl, pentenyl,isopentenyl, pentadienyl, hexenyl, isohexenyl, hexadienyl, heptenyl,octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl,tetradecenyl and pentadecenyl. Examples of a preferable aspect of the“lower alkenyl” include vinyl, allyl, propenyl, isopropenyl and butenyl.

In the present specification, the “lower alkynyl” includes straight orbranched alkynyl having a carbon number of 2 to 10, preferably a carbonnumber of 2 to 8, further preferably a carbon number of 3 to 6, havingone or more triple bonds at arbitrary positions. Specifically, examplesthereof include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl,octynyl, nonynyl and decynyl. These may further have a double bond at anarbitrary position. Examples of a preferable aspect of the “loweralkynyl” include ethynyl, propynyl, butynyl and pentynyl.

In the present specification, lower alkyl parts of the “lower alkyloxy”,the “lower alkylcarbonyl”, the “lower alkyloxycarbonyl”, the“carbocyclyl lower alkyl”, the “heterocyclyl lower alkyl”, the“carbocyclyloxy lower alkyl”, the “heterocyclyloxy lower alkyl”, the“halogeno lower alkyl”, the “carbocyclyl lower alkyloxy”, the“heterocyclyl lower alkyloxy”, the “carbocyclyl lower alkylthio”, the“heterocyclyl lower alkylthio”, the “carbocyclyl lower alkylamino”, the“heterocyclyl lower alkylamino”, the “halogeno lower alkyloxy”, the“lower alkyloxy lower alkyl”, the “lower alkyloxy lower alkyloxy”, the“lower alkylcarbonyl”, the “lower alkyloxycarbonyl”, the “loweralkylamino”, the “lower alkylcarbonylamino”, the “loweralkylaminocarbonyl”, the “lower alkyloxycarbonylamino”, the “loweralkylsulfonyl” and the “lower alkylsulfonylamino” are also the same asthe “lower alkyl”.

In the present specification, a lower alkenyl part of the “loweralkenylamino” is also the same as the “lower alkenyl”.

In the present specification, a lower alkynyl part of the “lower alkynylamino” is also the same as the “lower alkynyl”.

In the present specification, halogen parts of the “halogeno loweralkyl” and the “halogeno lower alkyloxy” are the same as theaforementioned “halogen”. Herein, arbitrary positions on an alkyl groupof the “lower alkyl” and the “lower alkyloxy” may be substituted by sameor different one or a plurality of halogen atoms, respectively.

In the present specification, the “carbocyclyl” means carbocyclyl havinga carbon number of 3 to 20, preferably a carbon number of 3 to 16, morepreferably a carbon number of 4 to 12, and includes cycloalkyl,cycloalkenyl, aryl and a non-aromatic fused carbocyclyl.

Specifically, the “cycloalkyl” is carbocyclyl having 3 to 16 carbonatoms, preferably 3 to 12 carbon atoms, more preferably 4 to 8 carbonatoms, and examples thereof include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl andcyclodecyl.

Specifically, the “cycloalkenyl” includes cycloalkenyl having one ormore double bonds at arbitrary positions in a ring of the cycloalkyl,and examples thereof include cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptynyl, cyclooctynyl and cyclohexadienyl.

Specifically, the “aryl” includes phenyl, naphthyl, anthryl andphenanthryl and, particularly, phenyl is preferable.

Specifically, the “non-aromatic condensed carbocyclyl” includes a groupin which two or more cyclic groups selected from the “cycloalkyl”, the“cycloalkenyl” and the “aryl” are condensed, and examples thereofinclude indanyl, indenyl, tetrahydronaphthyl, fluorenyl, adamantyl and agroup shown below.

Examples of a preferable aspect of the “carbocyclyl” include cycloalkyl,aryl and a non-aromatic fused carbocyclyl and, specifically, examplesthereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl and phenyl.

Carbocyclyl parts of the “carbocyclyl lower alkyl”, the “carbocyclyllower alkyloxy”, the “carbocyclyl lower alkylthio”, the “carbocyclyllower alkylamino”, the “carbocyclyloxy”, the “carbocyclylcarbonyl” andthe “carbocyclylaminocarbonyl” are as the same as the “carbocyclyl”.

In the present specification, the “heterocyclyl” includes heterocyclylsuch as heteroaryl, non-aromatic heterocyclyl, bicyclic condensedheterocyclyl and tricyclic condensed heterocyclyl having one or moresame or different hetero atoms arbitrarily selected from O, S and N in aring.

Specifically, examples of the “heteroaryl” include 5 to 6-memberedaromatic cyclic groups such as pyrrolyl, imidazolyl, pyrazolyl, pyridyl,pyridazinyl, pyrimidinyl, pyrazinyl, triazolyl, triazinyl, tetrazolyl,furyl, thienyl, isooxazolyl, oxazolyl, oxadiazolyl, isothiazolyl,thiazolyl and thiadiazolyl.

Specifically, examples of the “non-aromatic heterocyclyl” includedioxanyl, thiiranyl, oxyranyl, oxetanyl, oxathiolanyl, azetidinyl,thianyl, thiazolidinyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl,imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl,morpholinyl, morpholino, thiomorpholinyl, thiomorpholino,dihydropyridyl, tetrahydropyridyl, tetrahydrofuryl, tetrahydropyranyl,dihydrothiazolyl, tetrahydrothiazolyl, tetrahydroisothiazolyl,dihydrooxazinyl, hexahydroazepinyl, tetrahydrodiazepinyl,tetrahydropyridazinyl, hexahydropyrimidinyl and dioxolanyl.

Specific examples of the “bicyclic fused heterocyclyl” include indolyl,isoindolyl, indazolyl, indolizinyl, indolinyl, isoindolinyl, quinolyl,isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, naphthyridinyl,quinoxalinyl, purinyl, pteridinyl, benzopyranyl, benzimidazolyl,benzotriazolyl, benzisoxazolyl, be nzoxazolyl, benzoxadiazolyl, benzoisothiazolyl, benzothiazolyl, benzothiadiazolyl, benzofuryl,isobenzofuryl, benzothienyl, benzotriazolyl, thienopyridyl,thienopyrrolyl, thienopyrazolyl, thienopyrazinyl, furopyrrolyl,thienothienyl, imidazopyridyl, pyrazolopyridyl, thiazolopyridyl,pyrazolopyrimidinyl, pyrazolotriazinyl, pyridazolopyridyl,triazolopyridyl, imidazothiazolyl, pyrazinopyridazinyl, quinazolinyl,quinolyl, isoquinolyl, naphthyridinyl, dihydrothiazolopyrimidinyl,tetrahydroquinolyl, tetrahydroisoquinolyl, dihydrobenzofuryl,dihydrobenzoxazinyl, dihydrobenzimidazolyl, tetrahydrobenzothienyl,tetrahydrobenzofuryl, be nzodioxolyl, be nzodioxonyl, chromanyl,chromenyl, octahydrochromenyl, dihydrobenzodioxynyl,dihydrobenzoxezinyl, dihydrobenzodioxepinyl and dihydrothienodioxynyl.

Specific examples of the “tricyclic condensed heterocyclyl” includecarbazolyl, acridinyl, xanthenyl, phenothiazinyl, phenoxathiinyl,phenoxazinyl, dibenzofuryl, imidazoquinolyl, tetrahydrocarbazolyl, and agroup shown below.

Examples of a preferable aspect of the “heterecyclyl” include 5 to6-membered heteroaryl or non-aromatic heterocyclyl, and tricyclic fusedheterocyclyl.

Heterocyclyl parts of the “heterocyclyl lower alkyl”, the “heterocyclyllower alkyloxy”, the “heterocyclyl lower alkylthio”, the “heterocyclyllower alkylamino”, the “heterocyclyloxy”, the “heterocyclylcarbonyl” andthe “heterocyclylaminocarbonyl” are also the same as the “heterocyclyl”.

“Step B and Step C are performed continuously” refers to implementationof Step C after implementation of Step B without performing isolationoperation and column chromatography purification of a product producedin Step B. A reaction container for performing Step B may be the same asor different from a reaction container for performing Step C.

The “lower alkyl optionally substituted by substituent E” means that the“lower alkyl” is unsubstituted, or one or a plurality of chemicallyacceptable substituents selected from substituent E are bound thereto.When a plurality of substituents are bound, the plurality ofsubstituents may be the same or different. Examples thereof includemethyl, fluoromethyl, trifluoromethyl, chlorodifluoromethyl and

The “carbocyclyl optionally substituted by substituent E” means that the“carbocyclyl” is unsubstituted, or one or a plurality of chemicallyacceptable substituents selected from substituent E are bound thereto.When a plurality of substituents are bound, the plurality ofsubstituents may be the same or different. Examples thereof includefluorophenyl, difluorophenyl and methoxyfluorophenyl.

The “carbocyclyl lower alkyl optionally substituted by substituent E”means that the “carbocyclyl” and/or the “lower alkyl” are unsubstituted,or one or a plurality of chemically acceptable substituents selectedfrom substituent E are bound. When a plurality of substituents arebound, the plurality of substituents may be the same or different.Examples thereof include 4-fluorobenzyl, 2,4-difluorobenzyl,4-methoxy-2-fluorobenzyl and 4-methoxyphenyldifluoromethyl.

The “lower alkyloxy optionally substituted by substituent E”, the“carbocyclyl lower alkyloxy optionally substituted by substituent E”,the “heterocyclyl lower alkyloxy optionally substituted by substituentE”, the “lower alkenyl optionally substituted by substituent E”, the“amino optionally substituted by substituent E”, the “lower alkylaminooptionally substituted by substituent E”, the “lower alkenylaminooptionally substituted by substituent E”, the “lower alkynylaminooptionally substituted by substituent E”, the “carbocyclyl loweralkylamino optionally substituted by substituent E”, and the“heterocyclyl lower alkylamino optionally substituted by substituent E”have the same meaning.

The “carbocyclyl optionally substituted by substituent F” means that the“carbocyclyl” is unsubstituted, or one or a plurality of chemicallyacceptable substituents selected from substituent F are bound thereto.When a plurality of substituents are bound, the plurality ofsubstituents may be the same or different. Examples thereof includefluorophenyl, difluorophenyl and methoxyfluorophenyl.

The “lower alkyloxy optionally substituted by substituent F” means thatthe “carbocyclyl” part is unsubstituted, or one or a plurality ofchemically acceptable substituents selected from substituent F are boundthereto. When a plurality of substituents are bound, the plurality ofsubstituents may be the same or different. Examples thereof includefluorobenzyloxy, difluorobenzyloxy and methoxyfluorobenzyloxy.

The “heterocyclyl optionally substituted by substituent F”, the“heterocyclyl lower alkyloxy optionally substituted by substituent F”,the “carbocyclyl lower alkylthio optionally substituted by substituentF”, the “heterocyclyl lower alkylthio optionally substituted bysubstituent F”, the “carbocyclyl lower alkylamino optionally substitutedby substituent F”, the “heterocyclyl lower alkylamino optionallysubstituted by substituent F”, the “carbocyclyloxy optionallysubstituted by substituent F”, the “heterocyclyloxy optionallysubstituted by substituent F”, the “carbocyclylcarbonyl optionallysubstituted by substituent F”, the “heterocyclylcarbonyl optionallysubstituted by substituent F”, the “carbocyclylaminocarbonyl optionallysubstituted by substituent F” and the “heterocyclylaminocarbonyloptionally substituted by substituent F” have the same meaning.

“R^(3e)s of —N(R^(3e))₂ may be taken together to form a heterocycle”includes, for example, the following formulas.

Definition of the “heterocycle” in “R^(3e)s of —N(R^(3e))₂ may be takentogether to form a heterocycle” is also the same as described above.

The “amino protective groups” may be general protective groups for anamino group, and are exemplified as amino protective groups described,for example, in Protective Groups in Organic Synthesis, Theodora W Green(John Wiley & Sons). Preferable are a tert-butyloxycarbonyl group and abenzyloxycarbonyl group.

The “carboxyl protective groups” may be general protective groups for anamino group, and are exemplified as carboxyl protective groupsdescribed, for example, in Protective Groups in Organic Synthesis,Theodora W Green (John Wiley & Sons). Preferable examples thereofinclude a methyl group, an ethyl group, a tert-butyl group, amethoxymethyl group, an allyl group, a benzyl group and ap-methoxybenzyl group.

Examples of the “counter anion of ammonium cation” in X^(d) includehalogen, CH₃COO⁻, HCOO⁻, NO₃ ⁻, BF₄ ⁻, PF₆ ⁻, HO⁻, Ph-SO₃ ⁻, CH₃-Ph-SO₃⁻, CH₃—SO₃ ⁻, PO₄ ³⁻, SO₄ ²⁻ and HSO₄ ⁻. Preferable are halogen⁻,CH₃COO⁻, NO₃ ⁻ and SO₄ ²⁻. When the anion is divalent or trivalent, theNH₄ ⁺ cation indicates one not being charged state by binding of two orthree molecules, respectively. Specific examples of the NH₄ ⁺X^(d−)include (NH₄ ⁺)₂SO₄ ²⁻ and (NH₄ ⁺)₃PO₄ ³⁻.

The “leaving group” indicates a substituent which is left by anucleophilic reaction, and examples thereof include halogen, —O—SO₂—CH₃,—O—SO₂—CF₃, —O—SO₂-Ph and —O—SO₂-Ph-CH₃. Preferable is halogen.

The “derivative of formula (XA3)” indicates one in which an aldehydegroup or a carboxyl group is formed by oxidative cleavage of an olefinsite, or amide is formed by condensation reaction with a carboxyl group.Specifically, it is shown in the following formula (XA3a), formula(XA3b) or formula (XA3c):

(wherein R^(b) is hydrogen, lower alkyl optionally substituted bysubstituent E, lower alkenyl optionally substituted by substituent E,lower alkynyl optionally substituted by substituent E, carbocyclyl loweralkyl optionally substituted by substituent E, or heterocyclyl loweralkyl optionally substituted by substituent E, and other each symbol isdefined in Item 1 and Item 3). When the formula (XA3a) is reacted withthe formula (V3), the formula (XA4) in which R^(a) is hydrogen isproduced. When the formula (XA3b) is reacted with the formula (V3), theformula (XA4) in which R^(a) is hydroxy is produced. When the formula(XA3b) is reacted with the formula (V3), the formula (XA4) in whichR^(a) is hydroxy is produced. When the formula (XA3c) is reacted withthe formula (V3), the formula (XA4) in which R^(a) is lower alkylaminooptionally substituted by substituent E, lower alkenylamino optionallysubstituted by substituent E, lower alkynylamino optionally substitutedby substituent E, carbocyclyl lower alkylamino optionally substituted bysubstituent E, or heterocyclyl lower alkylamino optionally substitutedby substituent E is produced.

The “oxidative cleavage” in “an aldehyde group or a carboxyl group isformed by oxidative cleavage of an olefin site of the formula (XA3), oramide is formed by condensation reaction with a carboxyl group” can beperformed under generally known oxidation reaction condition (e.g. ozoneoxidation reaction, RuCl₃—NaIO₄ oxidation reaction etc.). When thereaction product is an aldehyde form (XA3a), a subsequent generaloxidation reaction condition (e.g. Cr₃-pyridine, PCC oxidation,SO₃-pyridine oxidation, NaClO₂ oxidation etc.) can derivatize the olefinsite into a carboxyl group.

The “amidation by a condensation reaction” can be performed by a generaldehydration condensation reaction (Mitsunobu reaction, reactions usingcarboxylic acid halide, carboxylic anhydride, or a condensing agent(e.g. WSC, carbonyldiimidazole, dicyclohexylcarbodiimide) etc.).

The “derivative of formula (XA4′)” indicates one in which an aldehydegroup or a carboxyl group is formed by oxidative cleavage of an olefinsite, or amide is formed by condensation reaction with a carboxyl group.Specifically, it is shown in the following formula (XA4′a), formula(XA4′b) or formula (XA4′c):

(wherein each symbol is defined in Item 1, and Item 3). When the formula(XA4′a) is reacted with the formula (V3′), the formula (XA4) in whichR^(a) is hydrogen is produced. When the formula (XA4′b) is reacted withthe formula (V3′), the formula (XA4) in which R^(a) is hydroxy isproduced. When the formula (XA4′c) is reacted with the formula (V3′),the formula (XA4) in which R^(a) is lower alkylamino optionallysubstituted by substituent E, lower alkenylamino optionally substitutedby substituent E, lower alkynylamino optionally substituted bysubstituent E, carbocyclyl lower alkylamino optionally substituted bysubstituent E, or heterocyclyl lower alkylamino optionally substitutedby substituent E is produced.

“An aldehyde group or a carboxyl group is formed by oxidative cleavageof an olefin site of formula (XA4′), or amide is formed by condensationreaction with a carboxyl group” is also the same as that of the case ofthe (XA3).

The production method according to the present invention will bedescribed below.

(Step A)

The present step is a step of reacting Compound (X1) and Compound (V1)to obtain a solution containing Compound (X2), as shown in the followingreaction formula.

Herein, the “solution” means a state where Compound (X2) is dissolved,and also includes one in a suspension state in which Compound (X2) isnot completely dissolved, but is dispersed, and one in a slurry state.Hereinafter, the “solution” in the present specification all similarlyincludes one in the suspension state or the slurry state.

(wherein each symbol is defined above)

Compound (X1) is a commercially available reagent, or can be obtained bya known method.

When R^(1d) is halogen, Compound (X1) in which R^(1d) is lower alkyloxyoptionally substituted by substituent E, carbocyclyl lower alkyloxyoptionally substituted by substituent E, heterocyclyl lower alkyloxyoptionally substituted by substituent E, or —OSi(R^(1e))₃ can be alsoobtained by adding an alcohol reagent such as a lower alcohol optionallysubstituted by substituent E, carbocyclyl lower alkyl alcohol optionallysubstituted by substituent E, heterocyclyl lower alkyl alcoholoptionally substituted by substituent E, or (R^(1e))₃Si—OH, andperforming a nucleophilic replacement reaction optionally in thepresence of a base, in a solvent.

Examples of the “lower alkyloxy optionally substituted by substituent E”in R^(1d) include methoxy, ethoxy, isopropoxy, trichloromethoxy andtrifluoromethoxy. Preferable is methoxy.

Examples of the “carbocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include benzyloxy, phenethyloxy,2,4-trifluorobenzyloxy and 4-methoxybenzyloxy. Preferable is benzyloxy.

Example of the “heterocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include pyridylmethyloxy.

A preferable aspect of R^(1d) is hydrogen, chloro, bromo, methoxy,benzyloxy or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(2d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include pyridylmethyl.

A preferable aspect of R^(2d) is methyl, ethyl, n-propyl, iso-propyl,tert-butyl, benzyl or the like.

A preferable aspect of R^(1e) is methyl, ethyl, n-propyl, iso-propyl,tert-butyl or the like.

As a reaction solvent of the nucleophilic replacement reaction forobtaining (X1), an aprotic solvent is preferable. Examples thereofinclude acetonitrile, tetrahydrofuran, dioxane, diethyl ether,dichloromethane, chloroform, toluene, xylene, ethyl acetate,N,N-dimethylformamide, N,N-dimethylacetamide anddimethylimidazolidinone.

The aforementioned base may be a base which can deprotonate an alcoholreagent, and examples thereof include n-butyllithium, tert-butyllithium,sodium-tert-butoxide, potassium-tert-butoxide, sodium-tert-pentoxide,sodium methoxide, sodium ethoxide, sodium hydride, lithiumdiisopropylamide and lithium bistrimethylsilylamide.

An amount of the base is about 1.0 to 3.0 molar equivalents relative toCompound (X1) in which R^(1d) is halogen.

An amount of the alcohol reagent is about 0.5 to 1.5 molar equivalentsrelative to Compound (X1) in which R^(1d) is halogen.

A reaction temperature is usually 0° C. to a refluxing temperature,preferably room temperature to 50° C.

A reaction time is usually 10 minutes to 50 hours, preferably 1 to 4hours.

Compound (V1) can be obtained as a commercially available reagent, orcan be obtained by a known method.

Examples of the “lower alkyl optionally substituted by substituent E” inP^(d) include methyl, ethyl and trifluoromethyl. A preferable aspect ofP^(d) is methyl.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3d) include methyl, ethyl and trifluoromethyl.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3e) include methyl, ethyl and trifluoromethyl.

A preferable aspect of R^(3d) is —N(CH₃)₂, —OCH₃, pyrrolidinyl or thelike.

Examples of a reaction solvent of a reaction for obtaining Compound (X2)by reacting Compound (X1) and Compound (V1) include acetonitrile,tetrahydrofuran, dioxane, diethyl ether, dichloromethane, chloroform,toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide and dimethylimidazolidinone.

An amount of Compound (V1) used is about 1.0 to 3.0 molar equivalentsrelative to Compound (X1), or Compound (V1) may be used as a solvent.

A reaction temperature is usually 0° C. to a refluxing temperature,preferably room temperature.

A reaction time is usually 30 minutes to 50 hours, preferably 2 to 8hours.

By the present step, a solution containing Compound (X2) is obtained.Compound (X2) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation.

(Step A′)

Alternatively, Compound (X2) can be also obtained by the followingreaction.

(wherein R^(7d) is halogen, lower alkyloxy optionally substituted bysubstituent E or —O—SO₂—R^(5e), and other each symbol is defined above)

Compound (Z1) is a commercially available reagent, or can be obtained bya known method.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(2d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include pyridylmethyl.

A preferable aspect of R^(2d) is methyl, ethyl, n-propyl, iso-propyl,tert-butyl, benzyl or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3d) include methyl, ethyl and trifluoromethyl.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3e) include methyl, ethyl and trifluoromethyl.

A preferable aspect of R^(3d) is —N(CH₃)₂, —OCH₃, pyrrolidinyl or thelike.

Examples of the “lower alkyloxy optionally substituted by substituent E”in R^(1d) include methoxy, ethoxy, isopropoxy, trichloromethoxy andtrifluoromethoxy. Preferable is methoxy.

Examples of the “carbocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include benzyloxy, phenethyloxy,2,4-trifluorobenzyloxy and 4-methoxybenzyloxy. Preferable is benzyloxy.

Example of the “heterocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include pyridylmethyloxy.

A preferable aspect of R^(1d) is hydrogen, chloro, bromo, methoxy,benzyloxy or the like.

Compound (Z1) is a commercially available reagent, or can be obtained bya known method

Examples of a preferable aspect of R^(7d) include chloro, bromo,methoxy, ethoxy, acetoxy, methanesulfonyloxy,trifluoromethanesulfonyloxy and paratoluenesulfonyloxy.

Examples of a reaction solvent of a reaction for obtaining Compound (X2)by reacting Compound (Z1) and Compound (Z2) include acetonitrile,tetrahydrofuran, dioxane, diethyl ether, dichloromethane, chloroform,toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide and dimethylimidazolidinone.

An amount of Compound (Z2) used is about 1.0 to 3.0 molar equivalentsrelative to Compound (Z1).

A reaction temperature is usually −10° C. to a refluxing temperature,preferably room temperature.

A reaction time is usually 10 minutes to 10 hours, preferably 1 to 4hours.

If necessary, a tertiary amine is added. Examples of the tertiary amineinclude pyridine, triethylamine, dimethylaminopyridine andN-methylmorpholine.

By the present step, a solution containing Compound (X2) is obtained.Compound (X2) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation.

(Step B)

The present step is a step of obtaining a solution containing Compound(X3), by reacting Compound (X2) and Compound (V2) optionally in thepresence of a base, as shown in the following reaction formula.

(wherein each symbol is defined above)

Examples of the “lower alkyloxy optionally substituted by substituent E”in R^(1d) include methoxy, ethoxy, isopropoxy, trichloromethoxy andtrifluoromethoxy. Preferable is methoxy.

Examples of the “carbocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include benzyloxy, phenethyloxy,2,4-trifluorobenzyloxy and 4-methoxybenzyloxy. Preferable is benzyloxy.

Example of the “heterocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include pyridylmethyloxy.

A preferable aspect of R^(1d) is hydrogen, chloro, bromo, methoxy,benzyloxy or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(2d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include pyridylmethyl.

A preferable aspect of R^(2d) is methyl, ethyl, n-propyl, iso-propyl,tert-butyl or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3d) include methyl, ethyl and trifluoromethyl.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(3e) include methyl, ethyl and trifluoromethyl.

A preferable aspect of R^(3d) is —N(CH₃)₂, —OCH₃, pyrrolidinyl or thelike.

Compound (V2) can be obtained as a commercially available reagent, orcan be obtained by a known method.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(4d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(4d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(4d) include pyridylmethyl.

Examples of a preferable aspect of R^(4d) include methyl, ethyl,n-propyl, iso-propyl, tert-butyl, benzyl and 4-methoxybenzyl.

Examples of a preferable aspect of R^(5d) include chloro, bromo,methoxy, ethoxy, acetoxy, methanesulfonyloxy,trifluoromethanesulfonyloxy and paratoluenesulfonyloxy. Particularly,chloro, methoxy and ethoxy are preferable.

Examples of a reaction solvent include acetonitrile, tetrahydrofuran,dioxane, toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine andN-methylpyrrolidinone.

Examples of the base include n-butyllithium, tert-butyllithium,sodium-tert-butoxide, potassium-tert-butoxide, sodium-tert-pentoxide,sodium methoxide, sodium ethoxide, sodium hydride, lithiumdiisopropylamide and lithium bistrimethylsilylamide.

An amount of the base used is about 1.0 to 5.0 molar equivalentsrelative to Compound (X2).

An amount of Compound (V2) used is about 1.5 to 5.0 molar equivalentsrelative to Compound (X2), or Compound (V2) may be used as a solvent.

A reaction temperature is usually −80° C. to a refluxing temperature,preferably −20° C. to 50° C.

A reaction time is usually 30 minutes to 50 hours, preferably 2 to 12hours.

By the present step, a solution containing Compound (X3) is obtained.Compound (X3) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

(Step B-II)

The present step is a step of obtaining a solution containing Compound(XA3) by reacting Compound (X2) and Compound (VA2) optionally in thepresence of a base, as shown in the following reaction formula.

(wherein each symbol is defined above)

Examples and preferable aspects of R^(1d), R^(2d), R^(3d) and R^(5d) inthe formulas (X2) and (VA2) are the same as those defined above,respectively.

Compound (VA2) can be obtained as a commercially available reagent, orby a known method.

Examples of the “lower alkyl optionally substituted by substituent E” ofR^(4e) include methyl and ethyl.

Examples of the “carbocyclyl optionally substituted by substituent E” ofR^(4e) include phenyl and cyclohexyl.

Examples of the “heterocyclyl optionally substituted by substituent E”of R^(4e) include pyridyl and piperazyl.

R^(4e) is preferably such that one is hydrogen, and the other is phenyl.

Examples of a reaction solvent include acetonitrile, tetrahydrofuran,dioxane, toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine andN-methylpyrrolidinone.

Examples of the base include n-butyllithium, tert-butyllithium,sodium-tert-butoxide, potassium-tert-butoxide, sodium-tert-pentoxide,sodium methoxide, sodium ethoxide, sodium hydride, lithiumdiisopropylamide and lithium bistrimethylsilylamide.

An amount of the base used is about 1.0 to 5.0 molar equivalentsrelative to Compound (XA2).

An amount of Compound (VA2) used is about 1.0 to 2.0 molar equivalentsrelative to Compound (X2).

A reaction temperature is usually −80° C. to 20° C., preferably −80° C.to −40° C. A reaction time is usually 5 minutes to 6 hours, preferably15 minutes to 2 hours.

By the present step, a solution containing Compound (XA3) is obtained.Compound (XA3) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

(Step C)

The present step is a step of obtaining Compound (X4) by reacting

Compound (X3) and Compound (V3), as shown in the following reactionformula.

(wherein each symbol is defined above)

Examples of the “lower alkyloxy optionally substituted by substituent E”in R^(1d) include methoxy, ethoxy, isopropoxy, trichloromethoxy andtrifluoromethoxy. Preferable is methoxy.

Examples of the “carbocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include benzyloxy, phenethyloxy,2,4-trifluorobenzyloxy and 4-methoxybenzyloxy. Preferable is benzyloxy.

Example of the “heterocyclyl lower alkyloxy optionally substituted bysubstituent E” in R^(1d) include pyridylmethyloxy.

A preferable aspect of R^(1d) is hydrogen, chloro, bromo, methoxy,benzyloxy or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(2d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(2d) include pyridylmethyl.

A preferable aspect of R^(2d) is methyl, ethyl, n-propyl, iso-propyl,tert-butyl or the like.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(4d) include methyl, ethyl, n-propyl, iso-propyl and tert-butyl.

Examples of the “carbocyclyl lower alkyl optionally substituted bysubstituent E” in R^(4d) include benzyl and 4-methoxybenzyl.

Examples of the “heterocyclyl lower alkyl optionally substituted bysubstituent E” in R^(4d) include pyridylmethyl.

Examples of a preferable aspect of R^(4d) include methyl, ethyl,n-propyl, iso-propyl, tert-butyl, benzyl and 4-methoxybenzyl.

Compound (V3) can be obtained as a commercially available reagent, orcan be obtained by a known method.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(6d) include HC(═O)—CH₂—, CH(—OH)₂—CH₂—, MeO—CH(—OH)—CH₂—,dimethoxyethyl, diethoxyethyl, CH₂═CH—CH₂—, HO—CH₂—CH(—OH)—CH₂—,

Examples of the “amino optionally substituted by substituent E” inR^(6d) include methylamino, ethylamino, benzyloxycarbonylamino andtert-butoxycarbonylamino.

Examples of the “carbocyclyl optionally substituted by substituent E” inR^(6d) include:

Examples of the “heterocyclyl optionally substituted by substituent E”in R^(6d) include:

Examples of a reaction solvent include acetonitrile, tetrahydrofuran,dioxane, toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine,N-methylpyrrolidinone, methanol, ethanol and isopropanol.

An amount of Compound (V3) used is about 1.0 to 2.0 molar equivalentsrelative to Compound (X3).

A reaction temperature is usually 0° C. to a refluxing temperature,preferably 20° C. to 70° C.

A reaction time is usually 30 minutes to 50 hours, preferably 2 to 12hours.

When R^(6d) of Compound (X4) produced is not a group having an aldehydegroup such as HC(═O)—CH₂—, MeO—CH(—OH)—CH₂— or CH(—OH₂)—CH₂— or theequivalent thereof, the group can be derivatized into such asHC(═O)—CH₂—, MeO—CH(—OH)—CH₂— or CH(—OH)₂—CH₂—, which is a group havingan aldehyde group or the equivalent thereof, by a method of deprotectinga protective group for an aldehyde group described in Protective Groupsin Organic Synthesis, Theodora W Green (John Wiley & Sons), or a knownmethod shown in International Publication No. 2006/116764 pamphlet, orInternational Publication No. 2006/088173 pamphlet.

For examples, when R^(6d) of Compound (X4) is dimethoxyethyl, it can bederivatized into HC(═O)—CH₂— by adding an acid to a solution containingCompound (X4). The acid is not particularly limited, and examplesthereof include hydrochloric acid, hydrobromic acid, sulfuric acid,nitric acid, phosphoric acid, p-toluenesulfonic acid, methanesulfonicacid, formic acid, acetic acid, trifluoroacetic acid, maleic acid andoxalic acid.

An amount of the acid used is 2.0 to 10.0 molar equivalents relative toCompound (X4). Acetic acid or formic acid may be used as a solvent, ormay be used by mixing with the aforementioned acid.

A reaction temperature is usually about 0° C. to 80° C., preferably 10°C. to 40° C.

A reaction time is usually 30 minutes to 50 hours, preferably 2 to 12hours.

When an amino group is protected with an amino protective group, adeprotected compound can be obtained by a method of deprotecting aprotective group for an amino group described in Protective Groups inOrganic Synthesis, Theodora W Green (John Wiley and Sons) or a knownmethod. An order of performing a deprotecting reaction can bearbitrarily changed.

By the present step, a solution containing Compound (X4) is obtained.Compound (X4) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

(Step C-II)

The present step is a step of obtaining a solution containing Compound(XA4) by reacting Compound (XA3) and Compound (V3) optionally in thepresence of a base, as shown in the following reaction formula.

(wherein each symbol is defined above)

Examples and preferable aspects of R^(1d), R^(2e) and R^(4e) in theformulas (XA3) and (VA4) are the same as those defined above,respectively.

Examples of the “lower alkylamino optionally substituted by substituentE” of R^(a) in the formula (XA4) include methylamino, ethylamino,isopropylamino, tert-butylamino, methoxymethylamino, methoxyethylamino,ethoxyethylamino, methoxypropylamino, hydroxyethylamino,piperazinylcarbonylethylamino, morpholinylcarbonylethylamino,methylaminoethylamino, methylsulfonylethylamino,tert-butylcarbonylaminoethylamino, isopropyloxycarbonylaminoethylamino,methylcarbonylaminoethylamino, aminoethylamino andtert-butyloxycarbonylaminoethylamino.

Examples of the “lower alkenylamino optionally substituted bysubstituent E” of R^(a) include ethylenylamino and propenylamino.

Examples of the “lower alkynylamino optionally substituted bysubstituent E” of R^(a) include propynylamino.

Examples of the “carbocyclyl lower alkylamino optionally substituted bysubstituent E” of R^(a) include benzylamino, difluorobenzylamino,chlorofluorobenzylamino, cyclopropylmethylamino, 4-fluorobenzylamino,cyclohexylmethylenylamino, cyclopropylamino, ethyloxycarbonylethylamino,carboxyethylamino, dimethylaminocarbonyl, 4-methoxybenzylamino and4-methylbenzylamino.

Examples of the “heterocyclyl lower alkylamino optionally substituted bysubstituent E” of R^(a) include pyridylmethylamino,tetrahydropyranylmethylenylamino and methylisoxazolylmethylenylamino.

Examples of the “lower alkyl optionally substituted by substituent E” inR^(6d) include HC(═O)—CH₂—, CH(—OH)₂—CH₂—, MeO—CH(—OH)—CH₂—,dimethoxymethyl, diethoxyethyl, CH₂═CH—CH₂—, HO—CH₂—CH(—OH)—CH₂—,

or, when R^(a) is hydroxy, examples of R^(6d) include:

or when R^(a) is the “lower alkylamino optionally substituted bysubstituent E”, the “lower alkenylamino optionally substituted bysubstituent E”, the “lower alkynylamino optionally substituted bysubstituent E”, the “carbocyclyl lower alkylamino optionally substitutedby substituent E”, or the “heterocyclyl lower alkylamino optionallysubstituted by substituent E”, examples of R^(6d) include:

Examples of the “amino optionally substituted by substituent E” inR^(6d) include methylamino, ethylamino, benzyloxycarbonylamino andtert-butoxycarbonylamino.

Examples of the “carbocyclyl optionally substituted by substituent E”R^(6d) include:

Examples of the “heterocyclyl optionally substituted by substituent E”in R^(6d) include:

Conversion from an olefin form (—CH═C(R^(4e))₂) to —CO—R^(a), when R^(a)is hydroxy, can be performed on aldehyde (R^(a) is hydrogen) obtainedunder generally known oxidation reaction condition (e.g. ozone oxidationreaction, RuCl₃—NaIO₄ oxidation reaction etc.), under subsequent generaloxidation reaction condition (e.g. Cr₃-pyridine, PCC oxidation,SO₃-pyridine oxidation, NaClO₂ oxidation etc.).

When R^(a) is the “lower alkylamino optionally substituted bysubstituent E”, the “lower alkenylamino optionally substituted bysubstituent E”, the “lower alkynylamino optionally substituted bysubstituent E”, the “carbocyclyl lower alkylamino optionally substitutedby substituent E”, or the “heterocyclyl lower alkylamino optionallysubstituted by substituent E”, the conversion can be performed on acarboxyl group (R^(a) is hydroxy) by a general dehydration condensationreaction (Mitsunobu reaction, reactions using carboxylic acid halide,carboxylic anhydride, or a condensing agent (e.g. WSC,carbonyldiimidazole, dicyclohexylcarbodiimide)).

Examples of a reaction solvent include acetonitrile, tetrahydrofuran,dioxane, toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine,N-methylpyrrolidinone, methanol, ethanol and isopropanol.

An amount of Compound (V3) used is about 1.0 to 3.0 molar equivalentsrelative to Compound (XA3).

A reaction temperature is usually 0° C. to a refluxing temperature,preferably 20° C. to 70° C.

A reaction time is usually 10 minutes to 50 hours, preferably 1 to 12hours.

When R^(6d) of Compound (X4) produced is not a group having an aldehydegroup such as HC(═O)—CH₂—, MeO—CH(—OH)—CH₂— or CH(—OH)₂—CH₂—, or theequivalent thereof, the group can be derivatized into such asHC(═O)—CH₂—, MeO—CH(—OH)—CH₂— or CH(—OH)₂—CH₂—, which is a group havingan aldehyde group or the equivalent thereof, by the aforementionedmethod.

When an amino group is protected with an amino protective group, adeprotected compound can be obtained by a method of deprotecting aprotective group for an amino group described in Protective Groups inOrganic Synthesis, Theodora W Green (John Wiley and Sons) or a knownmethod. An order of performing a deprotecting reaction can bearbitrarily changed.

By the present step, a solution containing Compound (XA4) is obtained.Compound (XA4) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

(Step B′) The present step is a step of obtaining Compound (X4′) byreacting Compound (X2) with Compound (V2) and Compound (V2′) optionallyin the presence of a base, as shown in following formula.

(wherein each symbol is defined above)

Examples and preferable aspects of R^(1d), R^(2d), R^(3d), R^(4d) andR^(5d) in the formulas (X2) and (V2) are the same as those describedabove, respectively.

Examples of a reaction solvent include acetonitrile, tetrahydrofuran,dioxane, toluene, xylene, ethyl acetate, N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine andN-methylpyrrolidinone.

Examples of the base include n-butyllithium, tert-butyllithium,sodium-tert-butoxide, potassium-tert-butoxide, sodium-tert-pentoxide,sodium methoxide, sodium ethoxide, sodium hydride, lithiumdiisopropylamide and lithium bistrimethylsilylamide.

An amount of the base used is about 1.0 to 5.0 molar equivalentsrelative to Compound (X2).

An amount of Compound (V2) used is about 1.0 to 3.0 molar equivalentsrelative to Compound (X2), or Compound (V2) may be used as a solvent.

A reaction temperature is usually −80° C. to a refluxing temperature,preferably −20° C. to 30° C.

A reaction time is usually 10 minutes to 10 hours, preferably 30 minutesto 4 hours.

Subsequently, Compound (V2′) is added to the reaction solution to allowto react.

Examples of Compound (V2′) include ammonium acetate, ammonium chloride,ammonium bromide, ammonium sulfate, ammonium hydrogen sulfate, ammoniumformate, ammonium nitrate, ammonium hydroxide, ammonium phosphate, NH₄⁺BF₄ ⁻, NH₄ ⁺PF₆ ⁻, NH₄ ⁺Ph-SO₃ ⁻, NH₄ ⁺CH₃-Ph-SO₃ ⁻ and NH₄ ⁺CH₃—SO₃ ⁻.Preferable are ammonium acetate, ammonium chloride, ammonium sulfate,ammonium hydrogen sulfate and ammonium formate

An amount of Compound (V2′) used is about 1.0 to 3.0 molar equivalentsrelative to Compound (X2).

A reaction temperature is usually 0° C. to a refluxing temperature,preferably 20° C. to 80° C.

A reaction time is usually 10 minutes to 10 hours, preferably 30 minutesto 4 hours.

By the present step, a solution containing Compound (X4′) is obtained.Compound (X4′) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

(Step C′)

The present step is a step of obtaining Compound (X4) by reactingCompound (X4′) and Compound (V3′) optionally in the presence of a base,as shown in following reaction formula.

(wherein each symbol is defined above)

Examples and preferable aspects of R^(1d), R^(2d), R^(4d) and R^(6d) inthe formulas (X4′) and (V3′) are the same as those defined above,respectively.

Examples of a “leaving group” in L^(d) include halogen, —O—SO₂—CH₃,—O—SO₂—CF₃, —O—SO₂-Ph or —O—SO₂-Ph-CH₃. Preferable is halogen

A method of derivatizing into an aldehyde group or the equivalentthereof when R^(6d) of Compound (X4) produced does not have an aldehydegroup such as HC(═O)—CH₂—, MeO—CH(—OH)—CH₂— or CH(—OH)₂—CH₂— or theequivalent thereof is the same as described above.

Examples of a reaction solvent include acetonitrile, ethyl acetate,N,N-dimethylformamide, N,N-dimethylacetamide, dimethylimidazolidinone,N-methylmorpholine and N-methylpyrrolidinone.

Examples of the base include potassium carbonate, cesium carbonate,sodium hydride, n-butyllithium, tert-butyllithium, sodium-tert-butoxide,potassium-tert-butoxide, sodium-tert-pentoxide, sodium methoxide,triethylamine, 4-dimethylaminopyridine, diisopropylethylamine and DBU(1,8-diazabicyclo[5.4.0]undec-7-ene).

An amount of the base used is about 1.0 to 5.0 molar equivalentsrelative to Compound (X4′).

An amount of Compound (V3′) used is about 1.0 to 4.0 molar equivalentsrelative to Compound (X4′), or Compound (V3′) may be used as a solvent.

A reaction temperature is usually 0° C. to a refluxing temperature,preferably 20° C. to 80° C.

A reaction time is usually 30 minutes to 24 hours, preferably 1 to 8hours.

By the present step, a solution containing Compound (X4) is obtained.Compound (X4) may be isolated by a general purification method(extraction, distillation, column chromatography, crystallization etc.),or may be used in a next reaction without isolation. Preferably, thecompound is isolated as a crystal from which impurities have beenremoved by crystallization.

When R^(6d) in Compound (X4) is —NH₂, Compound (X4) can be also obtainedby reacting an O-(2,4-dinitorophenyl)hydroxylamine reagent with Compound(X4′) optionally in the presence of a base.

Examples of a reaction solvent include N,N-dimethylformamide,N,N-dimethylacetamide, dimethylimidazolidinone, N-methylmorpholine andN-methylpyrrolidinone.

Examples of the base include potassium carbonate, cesium carbonate,sodium carbonate and lithium carbonate.

An amount of the base used is about 1.0 to 5.0 molar equivalentsrelative to Compound (X4′).

An amounts of the reagent used is about 1.0 to 4.0 molar equivalentsrelative to Compound (X4′).

A reaction temperature is usually 0° C. to a refluxing temperature,preferably 20° C. to 60° C.

A reaction time is usually 30 minutes to 24 hours, preferably 1 to 8hours.

(Step D-1)

The present step is a step of obtaining Compound (I), when Compound(X4A) has the following structure.

(wherein R¹⁵ is an amino protective group, hydrogen, lower alkyloptionally substituted by substituent E, lower alkenyl optionallysubstituted by substituent E, lower alkynyl optionally substituted bysubstituent E, carbocyclyl lower alkyl optionally substituted bysubstituent E, or heterocyclyl lower alkyl optionally substituted bysubstituent E, and each symbol is defined above)

A closed ring form can be obtained by generating an intramoleculardehydration condensation reaction (e.g. Mitsunobu reaction) on an amidosite (—CONH—R¹⁵) and hydroxy group of Compound (XA4). When R¹⁵ is anamino protective group, the intramolecular dehydration condensationreaction can be performed after the group is subjected to a knowndeprotecting reaction.

When R⁷ is not hydrogen, by performing a known nucleophilic replacementreaction on an amino group, objective R⁷ can be obtained.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing the known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be also directlyderivatized into —COR¹ by an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1e))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2007), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473, 2007)are also exemplified as a reaction of converting halogen into a hydroxygroup. As described above, when R^(1d) of a raw material is halogen,since it becomes possible to be derivatized as it is, the number ofreaction steps is deleted, and this can construct a more advantageousindustrial production method, as compared with a method of carrying outa reaction of protecting and/or deprotecting an alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting with a halogenating agentsuch as N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

(Step D-2)

The present step is a step of obtaining Compound (II), when Compound(X4A) has the following structure.

(wherein each symbol is defined above)

A closed ring form can be obtained by generating an intramoleculardehydration condensation reaction (e.g. Mitsunobu reaction) on an amidosite (—CONH—R¹⁵) and hydroxy group of Compound (XA4). When R¹⁵ is anamino protective group, the intramolecular dehydration condensationreaction can be performed after the group is subjected to a knowndeprotecting reaction.

When R⁷ is not hydrogen, by performing a known nucleophilic substitutionreaction on an amino group, objective R⁷ can be obtained.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing a known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be directlyderivatized into —COR¹ by an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1e))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2007), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba_(a))/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473,2007) are also exemplified as a reaction of converting halogen into ahydroxy group. As described above, when R^(1d) of a raw material ishalogen, since it becomes possible to be derivatized as it is, thenumber of reaction steps is deleted, and this can construct a moreadvantageous industrial production method, as compared with a method ofperforming a reaction of protecting and/or deprotecting an alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting a halogenating agent suchas N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

(Step D-3)

The present step is a step of obtaining Compound (I), when Compound(X4A) has the following structure.

(wherein each symbol is defined above)

A closed ring form can be obtained by generating an intramoleculardehydration condensation reaction (e.g. Mitsunobu reaction, amidationreaction using a condensing agent etc.) on a carboxyl group (—COOH) andamino group of Compound (XA4). When R¹⁵ is an amino protective group,the intramolecular dehydration condensation reaction can be preformedafter the group is subjected to a known deprotecting reaction.

When R⁷ is not hydrogen, by performing a known nucleophilic replacementreaction on an amino group, objective R⁷ can be obtained.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing a known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be directlyderivatized into —COR¹ by performing an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1e))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2027), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473, 2007)are also exemplified as a reaction of converting halogen into a hydroxygroup. Alternatively, when R^(1d) of a raw material is halogen, since itbecomes possible to be derivatized as it is, the number of reaction stepis deleted, and this can construct a more advantageous industrialproduction method, as compared with a method of performing a reaction ofprotecting and/or deprotecting an alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting a halogenating agent suchas N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

(Step D-4)

The present step is a step of obtaining Compound (II), when Compound(X4A) has the following structure.

(wherein each symbol is defined above)

A closed ring form can be obtained by generating an intramoleculardehydration condensation reaction (e.g. Mitsunobu reaction, an amidationreaction using a condensing agent etc.) on a carboxyl group (—COOH) andamino group of Compound (XA4). When R¹⁵ is an amino protective group,the intramolecular dehydration condensation reaction is performed afterthe group is subjected to a known deprotecting reaction.

When R⁷ is not hydrogen, by performing a known nucleophilic replacementreaction on an amino group, objective R⁷ can be obtained.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing a known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be also directlyderivatized into —COR¹ by performing an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1e))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2007), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473, 2007)are also exemplified as a reaction of converting halogen into a hydroxygroup. As described above, when R^(1d) of a raw material is halogen,since it becomes possible to be derivatized as it is, the number ofreaction step is deleted, and this can construct a more advantageousindustrial production method, as compared with a method of performing areaction of protecting and/or deprotecting an alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting a halogenating agent suchas N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

(Step D-5)

The present step is a step of obtaining Compound (II), when Compound(XA4) has the following structure.

(wherein each symbol is defined above)

A closed ring form can be obtained by generating a condensation reactionof an amido site (—CONH—R¹⁵) and amino group (—NH—R³) of Compound (X4)with a compound having a carboxyl group (R⁵—CO—R⁶). Examples of theR⁵—CO—R⁶ include paraformaldehyde and, in this case, R⁵ and R⁶ arehydrogen. When the condensation reaction is performed, an acid is added,if necessary. Examples of the acid include acetic acid, formic acid andsulfuric acid.

When R¹⁵ is an amino protective group, the condensation reaction can beperformed after the group is subjected to a known deprotecting reaction.

When R⁷ is not hydrogen, by performing a known nucleophilic replacementreaction on an amino group, objective R⁷ can be obtained.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing a known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be also directlyderivatized into —COR¹ by performing an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1e))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2007), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473, 2007)are also exemplified as a reaction of converting halogen into a hydroxygroup. As described above, when R^(1d) of a raw material is halogen,since it becomes possible to be derivatized as it is, the number ofreaction step is deleted, and this can construct a more advantageousindustrial production method, as compared with a method of performing areaction of protecting and/or deprotecting an alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting a halogenating agent suchas N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

(Step D-6)

The present step is a step of obtaining Compound (III), when Compound(X4) has the following structure.

(wherein each symbol is defined above)

A tricyclic compound can be obtained by reacting an aldehyde site ofCompound (X4) with an amino group and —Y—H of Compound (V4). When Z is asingle bond, R¹⁴ and R¹⁰ may be taken together to form 4 to 8-memberedheterocyclyl optionally substituted with substituent E and, in thiscase, Compound (III) becomes a tetracyclic compound.

Examples of Compound (V4) include 3-aminobutanol.

When R^(2d) is not hydrogen, a —COOR^(2d) site can be derivatized into—COR¹ by performing a known reaction (ester hydrolysis reaction,deprotective reaction of carboxyl protective group, etc.) to convertR^(2d) into hydrogen, and performing a known reaction (dehydrationcondensation reaction etc.) with carbocyclyl lower alkylamine optionallysubstituted by substituent E, lower alkylalcohol optionally substitutedby substituent E etc. In addition, when R^(2d) is a lower alkyl groupsuch as a methyl group or an ethyl group, the site can be also directlyderivatized into —COR¹ by performing an aminolysis reaction.

When R^(1d) is lower alkyloxy optionally substituted by substituent E,carbocyclyl lower alkyloxy optionally substituted by substituent E,heterocyclyl lower alkyloxy optionally substituted by substituent E, or—OSi(R^(1c))₃, it can be derivatized into a hydroxy group by subjectingto a known hydroxy deprotective reaction.

When R^(1d) is halogen, it can be derivatized into a hydroxy group byreacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Alternatively, sodium hydride/water (Bioorganic MedicinalChemistry Letters, 17, 1713, 2007), potassiumhydroxide/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/di-tert-butylarylphosphine (Journal of the American ChemicalSociety, 128, 10694, 2006), potassium phosphate hydrate(K₃PO₄.H₂O)/tris(dibenzylideneacetone)dipalladium(Pd₂dba₃)/tri-tert-butylphosphine (Tetrahedron Letters, 48, 473, 2007)are also exemplified as a reaction of converting halogen into a hydroxygroup. As described above, when R^(1d) of a raw material is halogen,since it becomes possible to be derivatized as it is, the number ofreaction step is deleted, and this can construct a more advantageousindustrial production method, as compared with a method of performing areaction of protecting and/or deprotecting alcohol.

When R^(1d) is hydrogen, it can be also derivatized into a hydroxy groupby converting R^(1d) into halogen by reacting a halogenating agent suchas N-bromosuccinimide, N-chlorosuccinimide, sulfuryl chloride etc.,similarly reacting with potassium trimethylsilanolate or lithiumtrimethylsilanolate, and adding an aqueous solution of an inorganicacid. Therefore, depending on reactivity of reaction substrates, R^(1d)can be appropriately selected.

In addition, in the above step, an order of an each reaction can beappropriately changed.

The present invention will be described more detail hereinbelow by wayof Examples and Test Examples of the present invention, but the presentinvention is not limited thereto. Respective symbols used in Exampleshave the following meanings.

DMF: N,N-dimethylformamide DMA: N,N-dimethylacetamide NMP:N-methylpyrrolidone DMI: Dimethylimidazolidinone THF: Tetrahydrofuran

MS: Methane sulfonyl

Ts: Paratoluenesulfonyl Boc: Tert-butoxycarbonyl

DIBALH: Diisobutylaluminum hydrideWSC: N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide

HOBt: 1-Hydroxybenzotriazole

HATU: O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate

NBS; N-bromosuccinimide NCS: N-chlorosuccinimide

TEMPO: 2,2,6,6-Tetramethylpiperidine-1-oxyl radicalPDC: Pyridinium dichloromateDEAD: Diethyl azodicarboxylateDIAD: Diisopropyl azodicarboxylate

DMAP: 4-Dimethylaminopyridine

mCPBA: M-chloroperbenzoic acidDBU: 1,8-Diazabicyclo[5,4,0]-7-undecene

The synthetic method of the present application will be shown below asExamples.

Example 1

First Step

A solution of benzyl alcohol (1.00 g, 9.25 mmol) in THF (3 ml) was addedto a suspension of sodium tert-pentoxide (2.55 g, 23.2 mmol) in THF (4ml) at room temperature under a nitrogen atmosphere, and the mixture wasstirred at 40° C. for 2 hours. This reaction solution was cooled in anice bath, and a solution of Compound 1 A (1.53 g, 10.2 mmol) in THF (3ml) was added dropwise at 0 to 10° C. After the reaction solution wasstirred at room temperature for 2 hours, 2 N hydrochloric acid (15 ml)was added, followed by extraction with ethyl acetate two times. Thecombined extracts were washed sequentially with water, saturated sodiumbicarbonate water, water and saturated sodium chloride water, and thendried with anhydrous sodium sulfate. The solvent was distilled off, andthe resulting oil was purified by silica gel column chromatography(n-hexane-ethyl acetate 4:1, v/v) to obtain 1.89 g (yield 92%) ofCompound 1B as an oil product.

¹H-NMR (CDCl₃) δ: 3.56 (2H, s), 3.71 (3H, s), 4.14 (2H, s), 4.59 (2H,s), 7.27-7.42 (5H, m).

Second Step

Compound 1B (1.80 g, 8.1 mmol) was dissolved in 1,4-dioxane (18 mL),N,N-dimethylformamide dimethyl acetal (1.45 g, 12.2 mmol) was added, andthe mixture was stirred at room temperature for 6 hours. The reactionsolution was concentrated under reduced pressure, and the residue waspurified by silica gel column chromatography (n-hexane-ethyl acetate1:4, v/v) to obtain 1.77 g (yield 79%) of Compound 1C as an oil product.

¹H-NMR (CDCl₃) δ: 2.90 (3H, br), 3.25 (3H, br), 3.69 (3H, s), 4.45 (2H,s), 4.59 (2H, s), 7.24-7.40 (5H, m), 7.73 (s, 1H).

Third Step

Sodium tert-butoxide (2.55 g, 23.2 mmol), dimethyl oxalate (639 mg, 5.41mmol) and DMI (3 ml) were added to a three-neck flask under a nitrogenatmosphere, and a solution of Compound 1C (0.50 g, 1.80 mmol) in DMI (2ml) was added dropwise thereto at 25 to 30° C. After stirring at roomtemperature for 7 hours, 2N hydrochloric acid (10 ml) was added, and themixture was stirred at room temperature for 15 hours. The reactionsolution was extracted with ethyl acetate two times, and the combinedextracts were washed sequentially with water, saturated sodiumbicarbonate water, water and saturated sodium chloride water, and thendried with anhydrous sodium sulfate. The solvent was distilled off, andthe resulting residue was purified by silica gel column chromatography(n-hexane-ethyl acetate 2:1 to 1:1, v/v) to obtain 488 mg (yield 85%) ofCompound 1D as a white crystal.

¹H-NMR (CDCl₃) δ: 3.89 (3H, s), 3.93 (3H, s), 5.34 (2H, s), 7.32-7.40(3H, m), 7.45-7.49 (2H, m), 8.50 (1H, s).

Example 2

First Step

A solution of benzyl alcohol (0.66 g, 6.1 mmol) in DMI (3 ml) was addedto a suspension of sodium tert-pentoxide (1.67 g, 15.2 mmol) in DMI (4ml) at room temperature under a nitrogen atmosphere, and the mixture wasstirred at 40° C. for 2 hours. This reaction solution was cooled in anice bath, and a solution of Compound 2A (1.10 g, 6.68 mmol) in DMI (3ml) was added dropwise at 0 to 10° C. The reaction solution was stirredat 0 to 5° C. for 2 hours, and at room temperature for 3 hours, and 2Nhydrochloric acid (15 ml) was added, followed by extraction with ethylacetate two times. The combined extracts were washed sequentially withwater, saturated sodium bicarbonate water, water and saturated sodiumchloride water, and then dried with anhydrous sodium sulfate. Thesolvent was distilled off, and the resulting oil product was purified bysilica gel column chromatography (n-hexane-ethyl acetate 4:1, v/v) toobtain 1.29 g (yield 90%) of Compound 2B as an oil product.

¹H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.2 Hz), 3.54 (2H, s), 4.14 (2H, s),4.17 (2H, q, J=7.2 Hz), 4.59 (2H, s), 7.28-7.40 (5H, m).

Second Step

Compound 2B (9.73 g, 41.2 mmol) was dissolved in toluene (45 ml),N,N-dimethylformamide dimethyl acetal (7.36 g, 61.8 mmol) was added, andthe mixture was stirred at room temperature for 5 hours. Water was addedto the reaction solution, followed by extraction with ethyl acetate twotimes. The combined extracts were washed sequentially with water, andsaturated sodium chloride water, and then dried with anhydrous magnesiumsulfate. The solvent was distilled off, and the resulting oil productwas purified by silica gel column chromatography (n-hexane-ethyl acetate1:1 to 3:7, v/v) to obtain 7.90 g (yield 66%) of Compound 2C as an oilproduct.

¹H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.2 Hz), 2.95 (3H, br), 3.22 (3H, br),4.15 (2H, q, J=7.2 Hz), 4.45 (2H, s), 4.59 (2H, s), 7.22-7.40 (5H, m),7.73 (1H, s).

Third Step

Sodium tert-butoxide (495 mg, 5.15 mmol) and DMI (2 ml) were added to athree-neck flask under a nitrogen atmosphere, and dimethyl oxalate (608mg, 5.15 mmol) and a solution of Compound 2C (0.50 g, 1.72 mmol) in DMI(3 ml) was added dropwise at 25 to 30° C. After stirring at roomtemperature for 4 hours, 2N hydrochloric acid (10 ml) was added, and themixture was stirred at room temperature for 15 hours. The reactionsolution was extracted with toluene two times, and the combined extractswere washed sequentially with water, saturated sodium bicarbonate water,water and saturated sodium chloride water, and then dried with anhydroussodium sulfate. The solvent was distilled off, and the resulting residuewas purified by silica gel column chromatography (n-hexane-ethyl acetate2:1, v/v) to obtain 420 mg (yield 74%) of Compound 2D as a whitecrystal.

¹H-NMR (CDCl₃) δ: 1.39 (3H, t, J=7.2 Hz), 3.88 (3H, s), 4.39 (2H, q,J=7.2 Hz), 5.34 (2H, s), 7.30-7.41 (3H, m), 7.45-7.50 (2H, m), 8.48 (1H,s).

Example 3

First Step

N,N-dimethylformamide dimethyl acetal (4.9 ml, 36.5 mmol) was addeddropwise to Compound 3A (5.0 g, 30.4 mmol) at 0° C. under cooling. Afterstirring at 0° C. for 1 hour, 100 ml of ethyl acetate was added to thereaction solution, followed by washing with 0.5N hydrochloric acid (50ml). The aqueous layer was separated, and extracted with ethyl acetate(50 ml). The organic layers were combined, washed sequentially withsaturated sodium bicarbonate water and saturated sodium chloride water,and then dried with anhydrous sodium sulfate. The solvent was distilledoff, and the resulting residue was purified by silica gel columnchromatography (n-hexane-ethyl acetate 1:1 (v/v)→ethyl acetate) toobtain 4.49 g (yield 67%) of Compound 3B as an oil product.

¹H-NMR (CDCl₃) δ:1.32 (3H, t, J=7.1 Hz), 2.90 (3H, br s), 3.29 (3H, brs), 4.23 (2H, q, J=7.1 Hz), 4.54 (2H, s), 7.81 (1H, s).

Second Step

Lithium hexamethyldisilazide (1.0 M toluene solution, 49 ml, 49.0 mmol)was diluted with tetrahydrofuran (44 ml), a solution of Compound 3B(4.49 g, 20.4 mmol) in tetrahydrofuran (10 ml) was added dropwisethereto at −78° C. under cooling, and a solution of ethyl oxalylchloride (3.35 g, 24.5 mmol) in tetrahydrofuran (10 ml) was addeddropwise. After stirring at −78° C. for 2 hours, a temperature wasraised to 0° C. After 2N hydrochloric acid was added to the reactionsolution, and the mixture was stirred for 20 minutes, the solution wasextracted with ethyl acetate (200 ml×2), and the organic layer waswashed with saturated sodium bicarbonate water and saturated sodiumchloride water and then dried with anhydrous sodium sulfate. The solventwas distilled off, and the resulting residue was purified by silica gelcolumn chromatography (n-hexane-ethyl acetate 7:3→5:5→0:10 (v/v)) toobtain 1.77 g (yield 31%) of Compound 3C as a white solid.

¹H-NMR (CDCl₃) δ:1.36-1.46 (6H, m), 4.35-4.52 (8H, m), 8.53 (1H, s).

Third Step

Aminoacetaldehyde dimethyl acetal (0.13 ml, 1.20 mmol) was added to asolution of Compound 3C (300 mg, 1.09 mmol) in ethanol (6 ml) at 0° C.,and the mixture was stirred at 0° C. for 1 hour and 30 minutes, at roomtemperature for 18 hours and, then, at 60° C. for 4 hours. After thesolvent was distilled off from the reaction solvent under reducedpressure, the resulting residue was purified by silica gel columnchromatography (n-hexane-ethyl acetate 5:5→0:10 (v/v)) to obtain 252 mg(yield 64%) of Compound 3D as an oil product.

¹H-NMR (CDCl₃) δ: 1.36-1.47 (6H, m), 3.42 (6H, s), 3.90 (2H, d, J=5.2Hz), 4.37 (3H, q, J=7.2 Hz), 4.50 (2H, q, J=7.2 Hz), 8.16 (1H, s).

Fourth Step

To a solution of Compound 3D (1.02 g, 2.82 mmol) in formic acid (10 ml),62%-H₂SO₄ (892 mg, 5.64 mmol) was added and the mixture was stirred atroom temperature for 16 hours. Formic acid was distilled off underreduced pressure, methylene chloride was added to the residue, andsaturated sodium chloride water was added to adjust a pH to 6.6. Themethylene chloride layer was separated, and the aqueous layer wasextracted with methylene chloride. The methylene chloride layers werecombined, and dried with anhydrous sodium sulfate. The solvent wasdistilled off to obtain 531.8 mg of Compound 3E as a yellow oil product.

¹H-NMR (CDCl₃) δ: 1.28-1.49 (6H, m), 4.27-4.56 (4H, m), 4.84 (2H, s),8.10 (1H, s), 9.72 (1H, s).

Fifth Step

Methanol (0.20 ml, 5.0 mmol), (R)-3-amino-butan-1-ol (179 mg, 2.0 mmol)and acetic acid (0.096 ml, 1.70 mmol) were added to a solution ofCompound 3E (531 mg, 1.68 mmol) in toluene (5 ml), and the mixture washeated to reflux for 4 hours. The reaction solution was cooled to roomtemperature, diluted with chloroform, and then washed with saturatedsodium bicarbonate water, and the aqueous layer was extracted withchloroform. The chloroform layers were combined, washed with saturatedsodium chloride water, and then dried with anhydrous sodium sulfate. Thesolvent was distilled off, and the resulting residue was purified bysilica gel column chromatography (chloroform-methanol 100:0→90:10) toobtain 309.4 mg of Compound 3F as a brown oil product.

¹H-NMR (CDCl₃) δ: 1.40 (3H, t, J=7.1 Hz), 1.40 (3H, d, J=7.1 Hz),1.55-1.61 (1H, m), 2.19-2.27 (1H, m), 4.00 (1H, d, J=1.5 Hz), 4.03 (1H,d, J=2.5 Hz), 4.10 (1H, dd, J=13.2, 6.3 Hz), 4.26 (1H, dd, J=13.2, 3.8Hz), 4.38 (2H, q, J=7.1 Hz), 5.00-5.05 (1H, m), 5.31 (1H, dd, J=6.4, 3.9Hz), 8.10 (1H, s).

Sixth Step

Potassium trimethylsilanolate (333 mg, 2.34 mmol) was added to asolution of Compound 3F (159 mg, 0.47 mmol) in 1,2-dimethoxyethane (2ml), and the mixture was stirred at room temperature for 7 hours.1N-hydrochloric acid and saturated sodium chloride water were added tothe reaction solution, followed by extraction with chloroform. Thechloroform layers were combined, and dried with anhydrous sodiumsulfate. The solvent was distilled off to obtain 34.4 mg (yield 25%) ofCompound 3G as an orange powder.

¹H-NMR (CDCl₃) δ: 1.46 (3H, d, J=3.5 Hz), 1.58-1.65 (1H, m), 2.26-2.30(1H, m), 4.06-4.10 (2H, m), 4.31 (1H, dd, J=13.8, 5.6 Hz), 4.48 (1H, dd,J=13.6, 3.9 Hz), 5.03 (1H, t, J=6.4 Hz), 5.36 (1H, dd, J=5.5, 4.0 Hz),8.44 (1H, s), 12.80 (1H, s), 14.90 (1H, s).

Example 4

First Step

N,N-dimethylformamide dimethyl acetal (12.2 ml, 92.2 mmol) was addeddropwise to Compound 4A (10.0 g, 76.8 mmol) at 0° C. under cooling.After stirring at 0° C. for 1 hour and 30 minutes and, then, at roomtemperature for 2 hours and 30 minutes, 100 ml of ethyl acetate wasadded to the reaction solution, and the solvent was distilled off. Theresulting residue was purified by silica gel column chromatography(n-hexane-ethyl acetate 5:5→0:10 (v/v)) to obtain 12.45 g (yield 88%) ofCompound 4B as an oil product.

¹H-NMR (CDCl₃) δ:1.32 (3H, t, J=7.1 Hz), 2.33 (3H, s), 3.04 (6H, br s),4.23 (2H, q, J=7.2 Hz), 7.68 (1H, s).

Second Step

Lithium hexamethyldisilazide (1.0M toluene solution, 24 ml, 24.0 mmol)was diluted with tetrahydrofuran (20 ml), a solution of Compound 4B(1.85 g, 10.0 mmol) in tetrahydrofuran (5 ml) was added dropwise theretoat −78° C. under cooling, and a solution of ethyl oxalyl chloride (1.34ml, 12.0 mmol) in tetrahydrofuran (5 ml) was added dropwise. Afterstirring at −78° C. for 2 hours, 2N-hydrochloric acid was added to thereaction solution, and the mixture was stirred at room temperature for20 minutes. The solution was extracted with ethyl acetate, and theorganic layer was washed sequentially with saturated sodium bicarbonatewater and saturated sodium chloride water, and then dried with anhydroussodium sulfate. The solvent was distilled off, and the resulting residuewas purified by silica gel column chromatography (n-hexane-ethyl acetate75:25→455:5 (v/v)) to obtain 1.03 g (yield 43%) of Compound 4C as abrown oil product.

¹H-NMR (CDCl₃) δ:1.38 (3H, t, J=7.1 Hz), 1.42 (3H, t, J=7.4 Hz),4.33-4.47 (4H, m), 7.19 (1H, s), 8.54 (1H, s).

Third Step

Aminoacetaldehyde dimethyl acetal (0.34 ml, 3.11 mmol) was added to asolution of Compound 4C (680 mg, 2.83 mmol) in ethanol (6.8 ml) at 0°C., and it was allowed to stand at room temperature for 16 hours. Afterthe solvent was distilled off from the reaction solution under reducedpressure, the resulting residue was purified by silica gel columnchromatography (n-hexane-ethyl acetate 90:10 (v/v)) to obtain 875 mg(yield 94%) of Compound 4D as an oil product.

¹H-NMR (CDCl₃) δ:1.38 (3H, t, J=7.1 Hz), 1.39 (3H, t, J=7.1 Hz), 3.40(611, s), 4.33 (2H, d, J=4.7 Hz), 4.37 (4H, q, J=7.1 Hz), 4.49 (1H, t,J=4.7 Hz), 7.06 (1H, s), 8.17 (1H, s).

Fourth Step

N-bromosuccinimide (1.46 g, 8.18 mmol) was added to a solution ofCompound 4D (2.68 g, 8.18 mmol) in N,N-dimethylformamide (10 ml), andthe mixture was stirred at room temperature for 48 hours. Aftersaturated sodium bicarbonate water was added to the reaction solution,the solution was extracted with ethyl acetate, and the organic layer waswashed sequentially with water and saturated sodium chloride water, andthen dried with anhydrous sodium sulfate. The solvent was distilled off,and the resulting residue was purified by silica gel columnchromatography (n-hexane-ethyl acetate 90:10 (v/v)) to obtain 2.83 g(yield 85%) of Compound 4E as an oil product.

¹H-NMR (CDCl₃) δ: 1.41 (3H, t, J=7.1 Hz), 1.48 (3H, t, J=7.1 Hz), 3.42(6H, s), 3.90 (2H, d, J=5.0 Hz), 4.39 (2H, q, J=7.1 Hz), 4.53 (3H, q,J=14.3 Hz), 4.54 (3H, s), 4.57 (3H, t, J=5.4 Hz), 8.19 (1H, s).

Fifth Step

To a solution of Compound 4E (2.23 g, 5.49 mmol) in formic acid (15 ml),62%-H₂SO₄ (1.74 g, 10.98 mmol) was added and the mixture was stirred atroom temperature for 8 hours. A 0.5N-aqueous sodium hydroxide solution(120 ml) was added, followed by extraction with methylene chloride. Themethylene chloride layers were combined, washed with saturated sodiumchloride water, and then dried with anhydrous sodium sulfate. Thesolvent was distilled off to obtain 1.31 g of Compound 4F as a whitepowder.

¹H-NMR (CDCl₃) δ: 1.31-1.46 (6H, m), 4.33-4.48 (4H, m), 4.82 (2H, s),8.11 (1H, s), 9.71 (1H, s).

Sixth Step

Methanol (0.44 ml, 10.9 mmol), (R)-3-amino-butane-1-ol (389 mg, 4.36mmol) and acetic acid (0.21 ml, 3.64 mmol) were added to a solution ofCompound 4F (1.31 g, 3.64 mmol) in toluene (13 ml), and the mixture washeated to reflux for 3 hours. The reaction solution was cooled to roomtemperature, diluted with chloroform, and then washed with saturatedsodium bicarbonate water, and the aqueous layer was extracted withchloroform. The chloroform layers were combined, washed with saturatedsodium chloride water, and then dried with anhydrous sodium sulfate. Thesolvent was distilled off, and the resulting residue was purified bysilica gel column chromatography (chloroform-methanol 100:0→90:10) toobtain 1.58 g of Compound 4G as an oil product.

¹H-NMR (CDCl₃) δ: 1.40 (3H, d, J=5.7 Hz), 1.56-1.60 (1H, m), 2.19-2.24(1H, m), 3.99 (1H, d, J=2.0 Hz), 4.02 (1H, d, J=2.4 Hz), 4.11 (1H, dd,J=13.3, 6.7 Hz), 4.28 (1H, dd, J=13.3, 3.9 Hz), 4.36 (3H, q, J=7.1 Hz),4.49-4.56 (1H, m), 4.98-5.03 (1H, m), 5.34 (1H, dd, J=6.6, 3.8 Hz), 8.07(1H, s).

Seventh Step

Potassium trimethylsilanolate (249 mg, 1.95 mmol) was added to asolution of Compound 4G (300 mg, 0.78 mmol) in 1,2-dimethoxyethane (3ml), and the mixture was stirred at room temperature for 1 hour.Potassium trimethylsilanolate (249 mg, 1.95 mmol) was additionallyadded, and the mixture was further stirred at 60° C. for 1 hour.1N-hydrochloric acid and saturated sodium chloride water were added tothe reaction solution, followed by extraction with chloroform. Thechloroform layers were combined, and dried with anhydrous sodiumsulfate. The solvent was distilled off to obtain 100.3 mg (yield 43%) ofCompound 3G as a yellow powder.

¹H-NMR (CDCl₃) δ: 1.46 (3H, d, J=3.5 Hz), 1.58-1.65 (1H, m), 2.26-2.30(1H, m), 4.06-4.10 (2H, m), 4.31 (1H, dd, J=13.8, 5.6 Hz), 4.48 (1H, dd,J=13.6, 3.9 Hz), 5.03 (1H, t, J=6.4 Hz), 5.36 (1H, dd, J=5.5, 4.0 Hz),8.44 (1H, s), 12.80 (1H, s), 14.90 (1H, s).

Example 5

First Step

Compound 5A (598 mg, 4.09 mmol) and N,N-dimethylformamide dimethylacetal (488 mg, 4.09 mmol) were dissolved in toluene (1 ml), and themixture was stirred at room temperature for 11 hours. The solvent wasdistilled off from the reaction solution under reduced pressure, and theresulting residue (containing Compound 5B) was used in Second stepwithout purification.

Second Step

Sodium tert-butoxide (400 mg, 4.16 mmol) was suspended in dimethylimidazolidinone (5 ml), a solution of the crude product obtained inFirst step in dimethylimidazolidinone (5 ml) was added thereto, asolution of dimethyl oxalate (983 mg, 8.32 mmol) in THF (10 ml) wasadded dropwise, and the mixture was stirred at room temperature for 45minutes. The reaction solution was poured into 2N hydrochloricacid-methanol (20 ml), and the mixture was stirred at 0° C. for 20minutes. Water was added, the solution was extracted with ethyl acetate,and the organic layer was washed sequentially with water, saturatedsodium bicarbonate water, and saturated sodium chloride water, and driedwith anhydrous sodium sulfate. After the solvent was distilled off, theresulting residue was purified by silica gel column chromatography toobtain 222 mg (yield: 22% from 5A) of Compound 5C.

¹H-NMR (CDCl₃) δ:3.91 (3H, s), 3.97 (3H, s), 4.05 (3H, s), 8.50 (1H, s).

Example 6

First Step

Lithium hexamethyldisilazide (1.0M toluene solution, 12 ml, 12.0 mmol)was diluted with tetrahydrofuran (11 ml), a solution of Compound 6A(1.46 g, 5.0 mmol) in tetrahydrofuran (2 ml) was added dropwise theretoat −78° C. under cooling, and a solution of ethyl oxalyl chloride (0.67ml, 6.0 mmol) in tetrahydrofuran (2 ml) was added dropwise. Afterstirring at −78° C. for 2 hours, ammonium acetate (500 mg) and aceticacid (10 ml) were added to the reaction solution, and the mixture wasstirred at 65° C. for 1 hour and 30 minutes. Water was added to thereaction solution, the solvent was extracted with ethyl acetate, and theorganic layer was washed sequentially with water, and saturated sodiumbicarbonate water, and dried with anhydrous sodium sulfate. The solventwas distilled off, and the resulting residue was purified by silica gelcolumn chromatography (N-hexane-ethyl acetate 55:45→45:55 (v/v)) toobtain 505.1 mg of Compound 6B as a yellow solid. It was washed withisopropyl ether-hexane (1:2), and dried under reduced pressure to obtain416.8 mg (yield 24%) of Compound 6B as a yellow crystal.

¹H-NMR (CDCl₃) δ:1.35 (3H, t, J=7.1 Hz), 1.46 (3H, t, J=7.1 Hz), 4.40(2H, q, J=7.2 Hz), 4.50 (2H, q, J=7.1 Hz), 5.20 (2H, s), 7.33-7.41 (3H,m), 7.49-7.52 (2H, m), 8.76 (1H, s), 11.61 (1H, br s).

Second Step

Cesium carbonate (73.3 mg, 0.23 mmol) and bromoacetaldehyde dimethylacetal (38.0 mg, 0.23 mmol) were added to a solution of Compound 6B(51.8 mg, 0.15 mmol) in N,N-dimethylformamide (1 ml), and the mixturewas stirred at room temperature overnight. Cesium carbonate (73.3 mg,0.23 mmol) and bromoacetaldehyde dimethyl acetal (38.0 mg, 0.23 mmol)were further added, and the mixture was further stirred at 100° C. for20 minutes. After water was added to the reaction solution, the solutionwas extracted with ethyl acetate, and the organic layer was washedsequentially with water and saturated sodium chloride water, and driedwith anhydrous sodium sulfate. The solvent was distilled off, and theresulting residue was purified by silica gel column chromatography(n-hexane-ethyl acetate 50:50→30:70 (v/v)) to obtain 35.3 mg (yield 54%)of Compound 6C as a colorless oil product.

¹H-NMR (CDCl₃) δ:1.26 (3H, t, J=7.1 Hz), 1.40 (3H, t, J=7.1 Hz), 3.39(6H, s), 3.91 (2H, d, J=5.0 Hz), 4.29 (2H, q, J=7.1 Hz), 4.40 (2H, q,J=7.2 Hz), 4.50 (1H, t, J=5.0 Hz), 5.30 (2H, s), 7.31-7.37 (3H, m),7.43-7.46 (2H, m), 8.12 (1H, s).

Example 7

First Step

Compound 6A (291 mg, 1.0 mmol) and dimethyl oxalate (354 mg, 3.0 mmol)were dissolved in dimethylimidazolidinone (1.4 ml), sodium methoxide(28%-methanol solution, 0.30 ml, 1.5 mmol) was added thereto, and themixture was stirred at room temperature for 2 hours.1,3-Dioxolan-2-yl-methylamine (154 mg, 1.5 mmol) and acetic acid (0.29ml, 5.0 mmol) were added thereto, and the mixture was stirred at roomtemperature for 38 hours. Saturated sodium bicarbonate water was addedto the reaction solution, followed by extraction with ethyl acetate. Theethyl acetate layers were combined, washed sequentially with water andsaturated sodium chloride water, and then dried with anhydrous sodiumsulfate. After the solvent was distilled off, the resulting residue waspurified by silica gel column chromatography (hexane-ethyl acetate33:67→15:85) to obtain 294.8 mg (yield 70%) of Compound 6C′ as a paleyellow oil product.

¹H-NMR (CDCl₃) δ: 1.43 (3H, t, J=7.1 Hz), 3.73-3.75 (2H, m), 3.81 (3H,s), 3.82-3.85 (2H, m), 4.21 (2H, d, J=2.2 Hz), 4.42 (2H, q, J=7.1 Hz),5.14 (1H, t, J=2.3 Hz), 5.32 (2H, s), 7.34-7.37 (3H, m), 7.44-7.46 (2H,m), 8.14 (1H, s).

Example 8

First Step

Aminoacetaldehyde dimethyl acetal (7.80 mmol) was added to a solution ofCompound 8A (900 mg, 2.60 mmol) in ethanol (5 ml), and the mixture wasstirred at room temperature for 22 hours. Ethyl acetate (5 ml) and water(5 ml) were added to the reaction solution, followed by extraction withethyl acetate (5 ml). After the organic layer was washed with water (10ml), the solvent was distilled off, and the resulting residue waspurified by silica gel column chromatography (n-hexane-ethyl acetate2:1) to obtain 0.37 g (yield 33%) of Compound 6C as a colorless oilproduct.

¹H-NMR (CDCl₃) δ: 7.90 (1H, s), 7.45-7.43 (5H, m), 5.30 (2H, s), 4.51(1H, t, J=5.1 Hz), 4.40 (2H, q, J=7.1 Hz), 4.30 (2H, q, J=7.1 Hz), 3.91(2H, d, J=5.1 Hz), 3.46 (6H, s), 1.40 (3H, t, J=7.1 Hz), 1.26 (3H, t,J=7.1 Hz).

Second Step

To a solution of Compound 6C (433.5 mg, 1.0 mmol) in formic acid (4 ml),62%-H₂SO₄ (316 mg, 2.0 mmol) was added and the mixture was stirred atroom temperature for 3 hours. Methylene chloride was added to thereaction solution, the solution was washed with a 0.5N-aqueous sodiumhydroxide solution (12 ml), and the aqueous layer was extracted withmethylene chloride. The methylene chloride layers were combined, washedwith saturated sodium chloride water, and then dried with anhydroussodium sulfate. The solvent was distilled off to obtain 207.6 mg (yield51%) of Compound 8C as a yellow foam product.

¹H-NMR (CDCl₃) δ: 1.23 (3H, t, J=7.1 Hz), 1.42 (3H, t, J=7.1 Hz), 4.25(2H, q, J=7.2 Hz), 4.42 (2H, q, J=7.1 Hz), 4.79 (2H, s), 5.34 (2H, s),7.31-7.53 (5H, m), 8.05 (1H, s), 9.67 (1H, s).

Example 9

First Step

Compound 9C (291.3 mg, 10 mmol) was dissolved in DMI (1.4 mL) in atwo-neck flask under a nitrogen atmosphere, dimethyl oxalate (354.3 mg,3.0 mmol), and sodium methoxide (28%-methanol solution 0.3 mL, 1.5 mmol)were added, and the mixture was stirred at room temperature for 2 hours.

2-(Aminomethyl)-1,3-dioxane (154.7 mg, 1.5 mmol) and acetic acid (0.29mL, 5.0 mmol) were added thereto, and the mixture was stirred at roomtemperature for 5 hours. Ethyl acetate (50 mL) was added to the reactionsolution, and the solution was washed sequentially with water (20 mL), a10%-aqueous ammonium chloride solution (20 mL), water (20 mL) andsaturated sodium chloride water (20 mL), and dried with anhydrousmagnesium sulfate. The solvent was distilled off, and the resultingresidue was purified by silica gel column chromatography (n-hexane-ethylacetate 1:1→1:3, v/v) to obtain 99.0 mg (yield 25%) of Compound 9C′ as awhite crystal.

¹H-NMR (CDCl₃) δ: 8.14 (1H, s), 7.44-7.42 (5H, m), 5.29 (2H, s), 5.12(1H, s), 4.19 (2H, s), 3.93 (3H, s), 3.83-3.70 (2H, m), 3.83 (2H, s).

Example 10

First Step

Compound 1D (318.3 mg, 1.0 mmol) and acetic acid (0.023 mL, 0.4 mmol)were dissolved in toluene (2 mL), a solution of tert-butylcarbazate(132.3 mg, 1.0 mmol) in toluene (2 mL) was added dropwise at 65° C., andthe mixture was stirred at 65° C. for 5 hours. Saturated sodiumbicarbonate water was added to the reaction solution, followed byextraction with ethyl acetate. The organic layer was washed withsaturated sodium chloride water, and dried with anhydrous magnesiumsulfate. The solvent was distilled off, and the resulting residue waspurified by silica gel column chromatography (hexane:ethylacetate=50:50→33:67) to obtain 315.3 mg (yield 72.9%) of Compound 10B.

¹H-NMR (DMSO-d6) δ: 1.36 (9H, s), 3.84 (6H, s), 5.11 (2H, s), 7.34-7.38(5H, m), 8.33 (1H, s), 11.11 (1H, br s).

Second Step

Compound 10B (310.3 mg, 0.72 mmol) was added to tetrahydrofuran-methanol(2 mL-1 mL), a 1N-aqueous lithium hydroxide solution (1.44 mL, 1.44mmol) was added, and the mixture was stirred at room temperature for 2hours. This was cooled in an ice water bath, 2N-hydrochloric acid wasadded, and ethyl acetate and water were added. The ethyl acetate layerwas separated, and the aqueous layer was extracted with ethyl acetate.The ethyl acetate layers were combined, washed with saturated sodiumchloride water, and dried with anhydrous sodium sulfate. The solvent wasdistilled off to obtain 284.1 mg (yield 94.3%) of Compound 10C.

¹H-NMR (CDCl₃) δ: 1.43 (9H, s), 3.98 (3H, s), 5.34 (214, s), 7.36-7.37(5H, m), 8.44 (1H, s), 14.53 (1H, br s).

Example 11

First Step

A solution of Compound 11A (12.8 g, 89.4 mmol) and pyridine (8.50 g, 107mmol) in dichloromethane (90 mL) was cooled to 1 to 3° C., and asolution of benzyloxyacetyl chloride (19.8 g, 107 mmol) indichloromethane (90 mL) was added dropwise over 50 minutes while thesame temperature was retained. After the reaction solution was stirredat the same temperature for 30 minutes, a temperature was graduallyraised to 15° C. over 60 minutes, and ice water was added. Thedichloromethane layer was separated, and the aqueous layer was extractedwith dichloromethane once. The combined extracts were washed with waterthree times, washed with saturated sodium chloride water, and thendried.

The solvent was distilled off, and the resulting oil product waspurified by subjecting it to silica gel column chromatography. First,the oil product was eluted first with n-hexane and, then, withn-hexane-ethyl acetate (1:1, v/v). The objective fraction wasconcentrated to obtain 22.2 g of Compound 11B as an oil product.

¹H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.2 Hz), 2.90 (3H, brs), 3.24 (3H,brs), 4.15 (2H, q, J=7.2 Hz), 4.45 (2H, s), 4.58 (2H, s), 7.25-7.38 (5H,m), 7.72 (1H, s).

Second Step

A 1N lithium hexamethyldisilazane THF solution (4.29 ml, 4.29 mmol) wascooled to −78° C., and a solution of Compound 11B (500 mg, 1.72 mmol)and cinnamoyl chloride (343.2 mg, 2.06 mmol) in THF (4 ml) was addeddropwise over 3 minutes while the same temperature was retained. Afterthe reaction solution was stirred at the same temperature for 25minutes, 2N hydrochloric acid (10 ml) was added, and the mixture wasfurther stirred at room temperature for 10 minutes. To the reactionsolution was added ethyl acetate, the organic layer was separated, andthe aqueous layer was extracted with ethyl acetate three times. Thecombined extracts were dried with sodium sulfate. The solvent wasdistilled off, and the resulting oil product was purified by subjectingit to silica gel column chromatography. From a fraction eluting withn-hexane-ethyl acetate (1:1, v/v), 364.3 mg (yield 56%) of Compound 11Cwas obtained as a solid.

¹H-NMR (CDCl₃) δ: 1.40 (3H, t, J=7.2 Hz), 4.39 (2H, q, J=7.2 Hz), 5.27(211, s), 6.99 (1H, d, J=16.2 Hz), 7.23 (1H, d, J=16.2), 7.26-7.48 (10H,m), 8.45 (1H, s).

Third Step

Under a nitrogen atmosphere, a solution of sodium periodate (625.8 mg,2.93 mmol) and 96% sulfuric acid (287.4 mg, 2.93 mmol) in water (8 ml)was added dropwise to a solution of Compound 11C and ruthenium chloride(2.76 mg, 0.0133 mmol) in MeCN (5 ml) at room temperature over 10minutes. After the reaction solution was stirred at the same temperaturefor 5 minutes, ethyl acetate was added, the organic layer was separated,and the aqueous layer was extracted with ethyl acetate two times. Thecombined extracts were dried with sodium sulfate. The solvent wasdistilled off, and the resulting oil product was purified by subjectingit to silica gel column chromatography. From a fraction elutingn-hexane-ethyl acetate (1:1, v/v), 303.2 mg (yield 75%) of Compound 11Dwas obtained as an oil product.

¹H-NMR (CDCl₃) δ: 1.39 (3H, t, J=6.9 Hz), 4.40 (2H, q, J=6.9 Hz), 5.54(2H, s), 7.37 (5H, s), 8.48 (1H, s), 9.85 (1H, s).

Fourth Step

A solution of 96% sulfuric acid (421.7 mg, 4.30 mmol) and amidosululicacid (642.7 mg, 6.62 mmol) in water (10 ml) was added to a solution ofCompound 11D (1.00 g, 3.31 mmol) in MeCN (15 ml) at room temperature,the mixture was stirred, and a solution of sodium chlorite (388.9 mg,4.30 mmol) in water (10 ml) was added dropwise over 5 minutes while thesame temperature was retained. The reaction solution was stirred at sametemperature for 5 minutes, and saturated sodium chloride water wasadded, followed by extraction with ethyl acetate three times. Thecombined extracts were dried with sodium sulfate. The solvent wasdistilled off, and the resulting oil product was purified by subjectingit to silica gel column chromatography. The column was eluted initiallywith chloroform and, then, chloroform-MeOH (7:3, v/v). When theobjective fraction was concentrated, 748.8 mg (yield 71%) of Compound11E was obtained as an oil product.

¹H-NMR (CDCl₃) δ: 1.40 (3H, t, J=7.2 Hz), 3.93 (1H, br s), 4.40 (2H, q,J=7.2 Hz), 5.61 (2H, s), 7.38-7.44 (10H, m), 8.52 (1H, s).

Fifth Step

WSC HCl (1.20 g, 6.28 mmol) and HOBt (551.6 mg, 4.08 mmol) were added toa solution of Compound 11E (1.00 g, 3.14 mmol) in DMF (10 ml) at roomtemperature, and the mixture was stirred at the same temperature for 90minutes. The reaction solution was cooled to 0° C., and a solution of2-methoxyethanamine (236.0 mg, 3.14 mmol) in DMF (2 ml) was addeddropwise over 3 minutes. The reaction solution was stirred at the sametemperature for 1 hour, and water was added, followed by extraction withethyl acetate three times. The extract was washed with water threetimes, and dried with sodium sulfate. The solvent was distilled off, andthe resulting oil product was subjected to silica gel chromatography topurify the oil product. The column was eluted initially withn-hexane-ethyl acetate (1:1, v/v) and, then, n-hexane-ethyl acetate(1:9, v/v). When the objective fraction is concentrated, 928.5 mg (yield79%) of Compound 11F was obtained as an oil product.

¹H-NMR (CDCl₃) δ: 1.39 (3H, t, J=7.2 Hz), 3.29 (3H, s), 3.41 (2H, t,J=5.4 Hz), 3.47-3.53 (2H, m), 4.39 (2H, q, J=7.2 Hz), 5.44 (2H, s), 7.36(3H, m), 7.44-7.47 (2H, m), 8.07 (1H, br s), 8.54 (1H, s).

Sixth Step

A solution of Compound 11F (500 mg, 1.33 mmol) and(S)-2-amino-3-phenylpropan-1-ol (604.2 mg, 4.0 mmol) in xylene (2 ml)was heated to 120° C., and the solution was stirred for 30 minutes. Thereaction solution was cooled to room temperature, the solvent wasdistilled off, and the resulting oil product was subjected to silica gelchromatography to purify the oil product. The column was elutedinitially with chloroform and, then, chloroform-MeOH (9:1, v/v). Whenthe objective fraction was concentrated, 487 mg (yield 72%) of Compound11G was obtained as an oil product.

¹H-NMR (CDCl₃) δ: 1.41 (3H, t, J=6.9 Hz), 2.24-2.34 (1H, m), 2.24-3.00(1H, m), 3.03-3.16 (1H, m), 3.05 (3H, m), 3.25-3.32 (2H, m), 4.13-4.19(1H, m), 4.17-4.30 (1H, m), 4.36-4.47 (1H, m), 4.51-4.54 (1H, m), 4.55(1H, d, J=10.5 Hz), 5.78 (1H, t, J=6.9 Hz), 7.17-7.26 (4H, m), 7.28-7.35(5H, m), 7.49 (1H, t, J=5.4 Hz), 6.32 (1H, s).

Seventh Step

A DEAD 40 wt % toluene solution (3.68 g, 8.45 mmol) was added dropwiseto a solution of Compound 11G (2.86 g, 5.63 mmol) and triphenylphosphine(2.21 g, 8.45 mmol) in THF (6 ml) at room temperature over 3 minutes.The reaction solution was stirred at the same temperature for 30minutes, the solvent was distilled off, and the resulting oil productwas subjected to silica gel chromatography to purify the oil product.From a fraction eluting with ethyl acetate-MeOH (9:1, v/v), 1.37 g(yield 50%) of Compound 11H was obtained as an oil product.

¹H-NMR (CDCl₃) δ: 1.31 (3H, t, J=7.2 Hz), 3.07 (2H, d, J=6.9 Hz), 3.33(3H, s), 3.57-3.80 (4H, m), 3.95 (1H, dd, J=3.0 Hz, 6.6 Hz), 4.01-4.14(1H, m), 4.16-4.34 (2H, m), 5.24 (1H, d, J=9.9 Hz), 5.51 (1H, d, J=9.9Hz), 7.01-7.03 (2H, m), 7.21-7.37 (5H, m), 7.41-7.58 (1H, m), 7.64-7.69(2H, m).

Eighth Step

A 2N aqueous sodium hydroxide solution (6 ml) was added to a solution ofCompound 11H (1.0 g, 2.04 mmol) in EtOH (6 ml), and the mixture wasstirred at room temperature for 30 minutes. The reaction solution wasneutralized with 2N hydrochloric acid, and the precipitated solid wasfiltered off, and dried to obtain 754 mg (yield 80%) of Compound 11I.

¹H-NMR (CDCl₃) δ: 3.10 (2H, d, J=7.8 Hz), 3.33 (3H, s), 3.57-3.69 (4H,m), 3.82-3.90 (1H, m), 3.95 (1H, dd, J=3.3 Hz, 13.8 Hz), 4.36 (1H, dd,J=6.3 Hz, 7.5 Hz), 5.36 (1H, d, J=10.2 Hz), 5.45 (1H, d, J=10.2 Hz),6.98-7.01 (2H, m), 7.28-7.39 (6H, m), 7.59 (2H, dd, J=1.8 Hz, 8.1 Hz),7.87 (1H, s).

Ninth Step

Compound 11I (1.0 g, 2.16 mmol) was dissolved in THF (10 ml), 10% Pd—C(200 mg) was added, and the mixture was subjected to a catalyticreduction reaction under a hydrogen stream. The catalyst was removed byfiltration, and the filtrate was concentrated. The resulting residue waswashed with ether to obtain 512 mg (yield 64%) of Compound 11.

¹H-NMR (CDCl₃) δ: 6.24 (2H, d, J=6.3 Hz), 3.36 (3H, s), 3.60-3.86 (5H,m), 4.14 (1H, d, J=12.9 Hz), 4.47 (1H, s), 7.03-7.05 (2H, m), 7.30-7.35(3H, m), 7.88 (1H, s), 12.68 (1H, s), 14.83 (1H, s).

Example 12

First Step

Compound 12A (1.53 g, 5.80 mmol) was dissolved in THF (6 ml) and water(6 ml), potassium carbonate (2.41 g, 17.4 mmol) was added, the mixturewas stirred, and benzyl chloroformate (1.09 g, 6.38 mmol) was addeddropwise at 0° C. After stirring at 0° C. for 10 minutes, the mixturewas stirred at room temperature for 2 hours. The reaction solution waspoured into an aqueous sodium bicarbonate solution, followed byextraction with ethyl acetate. The extract was washed with 1Nhydrochloric acid and saturated sodium chloride water, and dried withsodium sulfate. The solvent was distilled off to obtain 2.32 g ofCompound 12B as a colorless gummy substance.

¹H-NMR (CDCl₃) δ: 1.98 (1H, brs), 3.55 (1H, m), 3.75 (1H, m), 4.20 (1H,d, J=10.5 Hz), 4.58 (1H, m), 4.83 (1H, brs), 5.07 (2H, s), 7.16-7.39(15H, m).

Second Step

Compound 12B (1.94 g, 5.37 mmol), triphenylphosphine (2.11 g, 8.05 mmol)and phthalimide (948 mg, 6.44 mmol) were added to THF (20 ml), anddiisopropyl azodicarboxylate (2.2M in toluene, 3.66 ml, 8.05 mmol) wasadded dropwise at room temperature. After stirring at room temperaturefor 4 hours, the solvent was distilled off under reduced pressure. Theresulting crude product was purified by silica gel column chromatography(n-hexane-ethyl acetate, 1:1, v/v) to obtain 2.39 g of Compound 12C as acolorless solid.

¹H-NMR (CDCl₃) δ: 3.73 (2H, m), 4.05 (1H, d, J=10.1 Hz), 4.70 (1H, d,J=9.6 Hz), 4.77 (2H, d, J=7.2 Hz) 5.02 (1H, m), 7.03-7.42 (15H, m), 7.68(2H, dd, J=5.7, 2.1 Hz), 7.78 (2H, dd, J=5.7, 2.1 Hz).

Third Step

Compound 12C (2.39 g, 4.87 mmol) was added to THF (20 ml) and methanol(20 ml), hydrazine hydrate (4.88 g, 97.4 mmol) was added, and themixture was stirred at 50° C. for 4 hours. The white precipitate wasremoved by filtration, followed by washing with methanol. The filtratewas distilled off under reduced pressure, and the resulting crudeproduct was purified by amino column chromatography(chloroform-methanol, 99:1, v/v) to obtain 1.41 g of Compound 12D as acolorless solid.

¹H-NMR (CDCl₃) δ: 2.63 (1H, dd, J=13.2, 5.8 Hz), 2.86 (1H, d, J=9.9 Hz),4.07 (1H, d, J=10.4 Hz), 4.53 (1H, m), 4.81 (1H, m), 5.00 (2H, d, 8.4Hz), 7.20-7.36 (10H, m).

Fourth Step

Compound 12D (1.41 g, 3.91 mmol) was dissolved in THF (15 ml), and Boc₂O(896 mg, 4.11 mmol) was added at room temperature. After stirring for1.5 hours, the solvent was concentrated under reduced pressure. Theresulting crude product was purified by silica gel column chromatography(n-hexane-ethyl acetate, 1.1, v/v) to obtain 1.77 g of Compound 12E as acolorless solid.

¹H-NMR (CDCl₃) δ: 1.41 (9H, s), 3.23 (2H, brm), 3.97 (1H, d, J=9.8 Hz),4.58-4.80 (3H, m), 5.00 (2H, d, J=9.8 Hz), 7.15-7.29 (10H, m).

Fifth Step

Compound 12E (1.73 g, 3.76 mmol) and palladium active carbon (10%, wet,200 mg) were added to methanol (20 ml), and the mixture was stirred atroom temperature for 1 hour under a hydrogen atmosphere. Afterfiltration with Celite, the solvent was concentrated under reducedpressure to obtain 1.01 g of a colorless oily substance 12F.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 2.82 (1H, m), 3.31 (1H, m), 3.73 (2H, d,J=6.9 Hz), 4.98 (1H, s), 7.18-7.39 (10H, m).

Sixth Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (974 mg, 3.06mmol) and 12F (999 mg, 3.06 mmol) were added to toluene (10 ml), and themixture was stirred at 110° C. for 5 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 98:2,v/v) to obtain 1.51 g of Compound 12G as a pale yellow solid.

¹H-NMR (CDCl₃) δ: 1.36 (9H, s), 3.40 (1H, m), 3.53 (1H, m), 3.82 (3H,s), 3.91 (3H, s), 4.29 (1H, d, J=11.3 Hz), 4.78 (1H, m), 4.82 (1H, m),5.11 (1.9H, d, J=7.5 Hz), 7.10-7.38 (10H, m), 8.27 (1H, s).

Seventh Step

To Compound 12G (1.45 g, 2.31 mmol) was added 4N HCl (ethyl acetatesolution, 20 ml), and the mixture was stirred at room temperature for1.5 hours. After the solvent was diltilled off under reduced pressure,an aqueous saturated sodium bicarbonate solution was added, and themixture was stirred at room temperature for 1.5 hours. This wasextracted with chloroform, and dried with sodium sulfate. After thesolvent was diltilled off under reduced pressure, the resulting crudeproduct was purified by silica gel column chromatography(chloroform-methanol, 95.5, v/v) to obtain 1.01 g of Compound 12H as acolorless solid.

¹H-NMR (CDCl₃) δ: 3.40 (1H, dd, J=13.6, 6.6 Hz), 3.78 (3H, s), 3.80 (1H,m), 4.37 (1H, d, J=11.6 Hz), 4.59 (1H, d, J=11.0 Hz), 5.43 (2H, d,J=10.2 Hz), 5.93 (1H, d, J=5.8 Hz), 7.03-7.21 (5H, m), 7.37 (9H, m),7.63 (2H, m).

Eighth Step

Compound 12H (50 mg, 0.10 mmol) was dissolved in DMF (1 ml), and cesiumcarbonate (165 mg, 0.50 mmol) was added. After stirring at roomtemperature for 30 minutes, iodomethane (0.032 ml, 0.50 mmol) was added,and the mixture was stirred at room temperature for 3.5 hours. Thereaction solution was poured into water, and this was extracted withethyl acetate, and dried with sodium sulfate. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 95:5,v/v) to obtain 49 mg of Compound 121 as a colorless solid.

Ninth Step

Compound 121 (49 mg, 0.096 mmol) was dissolved in THF (0.5 ml) andmethanol (0.5 ml), a 2N aqueous sodium hydroxide solution (0.24 ml, 0.48mmol) was added at room temperature, and this was stirred as it was for1.5 hours. 1N hydrochloric acid was added, and this was extracted withethyl acetate, and dried with sodium sulfate. After the solvent wasdistilled off under reduced pressure, 54 mg of Compound 12J was obtainedas a colorless solid.

MS: m/z=481 [M+H]⁺.

Tenth Step

Trifluoroacetic acid (1 ml) was added to Compound 12J obtained in Ninthstep, and mixture was stirred at room temperature for 1 hour. Afterconcentration under reduced pressure, an aqueous sodium bicarbonatesolution and 2N hydrochloric acid were used to adjust a pH to 3, andthis was extracted with chloroform, and dried with sodium sulfate. Afterthe solvent was distilled off under reduced pressure,chloroform-methanol-ethyl ether were added, and the precipitated solidwas filtered off to obtain 26 mg of Compound 12 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 3.01 (3H, s), 3.26 (1H, t, J=14.4 Hz), 4.23 (1H, dd,J=13.5, 3.8 Hz), 4.57 (1H, d, J=11.6 Hz), 5.78 (1H, d, J=11.3 Hz),7.16-7.70 (10H, m), 8.00 (1H, s), 13.00 (1H, s), 15.10 (1H, s).

MS: m/z=405 [M+H]⁺.

Example 13

First Step

Compound 28A (3.20 g, 17.1 mmol) was added to THF (20 ml), triethylamine(2.60 ml, 18.8 mmol) was added, and the mixture was stirred at roomtemperature for 10 minutes. Boc₂O (4.09 g, 18.8 mmol) was added at roomtemperature, and the mixture was stirred as it was for 2 hours. Thesolvent was distilled off under reduced pressure, and water was added,followed by extraction with ethyl acetate. The organic layer was washedwith saturated sodium chloride water, and dried with sodium sulfate. Thesolvent was distilled off under reduced pressure to obtain 5.17 g ofCompound 28b as a colorless solid.

¹H-NMR (CDCl₃) δ: 1.52 (9H, s), 2.77 (2H, m), 3.03-3.12 (1H, m), 3.38(1H, m), 3.90-3.98 (1H, m), 4.93 (1H, brs), 7.20-7.35 (5H, m).

Second Step

Compound 28B (4.29 g, 17.1 mmol), triphenylphosphine (5.37 g, 20.5 mmol)and phthalimide (2.76 g, 18.8 mmol) were added to THF (60 ml), anddiethyl azodicarboxylate (2.2M in toluene, 11.6 ml, 25.6 mmol) was addeddropwise at room temperature. After stirring at room temperature for 1hour, the solvent was distilled off under reduced pressure. Theresulting crude product was purified by silica gel column chromatography(n-hexane-ethyl acetate, 2:1, v/v) to obtain 6.13 g of Compound 28C as acolorless solid.

¹H-NMR (CDCl₃) δ: 1.30 (9H, s), 3.14 (1H, dd, J=13.8, 6.2 Hz), 3.39 (2H,m), 3.87 (1H, m), 4.67 (1H, m), 4.81 (1H, brs), 7.16-7.19 (5H, m), 7.66(2H, dd, J=5.3, 3.1 Hz), 7.75 (2H, dd, J=5.7, 3.0 Hz).

Third Step

Compound 28C (1.00 g, 2.63 mmol) was added to THF (7 ml) and methanol (7ml), hydrazine hydrate (2.63 g, 52.6 mmol) was added, and the mixturewas stirred at 50° C. for 2 hours. The white precipitate was removed byfiltration, followed by washing with methanol. After the filtrate wasdistilled off under reduced pressure, the resulting crude product waspurified by amino column chromatography (chloroform-methanol, 99:1, v/v)to obtain 249 mg of Compound 28D as a colorless solid.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.95 (2H, brs), 2.55-3.31 (5H, m), 5.06(1H, brs), 7.18-7.33 (5H, m).

Fourth Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (313 mg, 0.983mmol) and 28D (246 mg, 0.983 mmol) were added to toluene (3 ml), and themixture was stirred at 100° C. for 2.5 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 98.2,v/v) to obtain 320 mg of Compound 28E as a pale yellow gummy substance.

¹H-NMR (CDCl₃) δ: 1.42 (9H, s), 3.07 (2H, m), 3.56 (2H, m), 3.68 (3H,s), 3.95 (3H, s), 4.26 (1H, s), 4.86 (1H, s), 5.18 (1H, d, J=10.8 Hz),5.22 (1H, d, J=10.8 Hz), 7.01 (2H, m), 7.24-7.38 (8H, m), 8.22 (1H, s).

MS: m/z=551 [M+H]⁺.

Fifth Step

To Compound 28E (315 mg, 0.572 mmol) was added to 4N HCl (ethyl acetatesolution, 5 ml), and the mixture was stirred at room temperature for 30minutes. The solvent was distilled off under reduced pressure, anaqueous saturated sodium bicarbonate solution was added, and this wasextracted with chloroform, and dried with sodium sulfate. After thesolvent was distilled off under reduced pressure, the resulting crudeproduct was purified by silica gel column chromatography(chloroform-methanol, 95:5, v/v) to obtain 210 mg of Compound 28F as acolorless solid.

¹H-NMR (CDCl₃) δ: 3.07-3.15 (2H, m), 3.34 (1H, dd, J=13.2, 6.0 Hz), 3.74(2H, m), 3.86 (3H, s), 4.12 (1H, m), 5.27 (1H, d, J=10.1 Hz), 5.47 (1H,d, J=10.1 Hz), 6.76 (1H, d, J=6.4 Hz), 7.04 (2H, m), 7.32 (6H, m), 7.62(2H, dd, J=7.7, 1.4 Hz), 7.70 (1H, s).

MS: m/z=419 [M+H]⁺.

Sixth Step

Compound 28F (50 mg, 0.12 mmol) was dissolved in DMF (1 ml), and cesiumcarbonate (195 mg, 0.597 mmol) was added After stirring at roomtemperature for 30 minutes, iodoethane (0.048 ml, 0.60 mmol) was added,and the mixture was stirred at room temperature for 3.5 hours. Thereaction solution was poured into water, and this was extracted withethyl acetate, and dried with sodium sulfate. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 95:5,v/v) to obtain 47 mg of Compound 28G as a colorless solid.

¹H-NMR (CDCl₃) δ: 1.22 (3H, t, J=7.2 Hz), 3.00-3.15 (2H, m), 3.28 (1H,dd, J=13.6, 1.6 Hz), 3.48 (1H, m), 3.75 (1H, m), 3.85 (3H, s), 3.88 (1H,dd, J=13.3, 3.2 Hz), 4.15 (1H, m), 5.25 (1H, d, J=9.9 Hz), 5.50 (1H, d,J=9.9 Hz), 7.04 (2H, m), 7.29-7.38 (6H, m), 7.60 (1H, s), 7.68 (2H, m).

MS: m/z=447 [M+H]⁺.

Seventh Step

Compound 28G (47 mg, 0.11 mmol) was dissolved in THF (0.5 ml) andmethanol (0.5 ml), a 2N aqueous sodium hydroxide solution (0.26 ml, 0.53mmol) was added at room temperature, and the mixture was stirred as itwas for 1 hour. 1N hydrochloric acid was added, and this was extractedwith ethyl acetate, and dried with sodium sulfate. The solvent wasdistilled off under reduced pressure to obtain 40 mg of Compound 28H asa colorless solid.

MS: m/z=433 [M+H]⁺.

Eighth Step

Trifluoroacetic acid (1 ml) was added to Compound 28H obtained inSeventh step, and the mixture was stirred at room temperature for 1hour. After concentration under reduced pressure, a pH was adjusted to 3with an aqueous sodium bicarbonate solution and 2N hydrochloric acid,and this was extracted with chloroform, and dried with sodium sulfate.After the solvent was distilled off under reduced pressure,chloroform-methanol-ethyl ether were added, and the precipitated solidwas filtered off to obtain 17 mg of Compound 28 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 1.17 (3H, t, J=7.2 Hz), 3.08 (2H, m), 3.51-3.63 (3H,m), 4.08 (1H, dd, J=13.6, 3.9 Hz), 5.03 (1H, brs), 7.21 (5H, m), 8.07(1H, s), 12.98 (1H, s), 15.07 (1H, brs).

MS: m/z=343 [M+H]⁺.

Example 14

First Step

Compound 43A (2.00 g, 6.11 mmol), triphenylphosphine (2.40 g, 9.16 mmol)and phthalimide (1.08 g, 7.33 mmol) were added to THF (20 ml), diethylazodicarboxylate (2.2M in toluene, 4.16 ml, 9.16 mmol) was addeddropwise at room temperature. After stirring at room temperature for 3hours, the solvent was distilled off under reduced pressure. Theresulting crude product was purified by silica gel column chromatography(n-hexane-ethyl acetate, 1:1, v/v) to obtain 2.39 g of Compound 43B as acolorless solid.

¹H-NMR (DMSO-d6) δ: 1.00 (9H, s), 3.30 (1H, m), 3.61 (1H, dd, J=13.4,10.2 Hz), 4.15 (1H, d, J=12.2 Hz), 4.75 (1H, m), 6.79 (1H, d, J=9.5 Hz),7.25 (15H, m), 7.76-7.89 (4H, m).

Second Step

Compound 43B (2.06 g, 4.51 mmol) was added to THF (20 ml) and methanol(20 ml), hydrazine hydrate (4.52 g, 90.2 mmol) was added, and themixture was stirred at 60° C. for 5 hours. The white precipitate wasremoved by filtration, followed by washing with methanol. After thefiltrate was distilled off under reduced pressure, the resulting crudeproduct was purified by amino column chromatography(chloroform-methanol, 99:1, v/v), n-hexane was added, and theprecipitated solid was filtered off to obtain 1.25 g of Compound 43C asa colorless solid.

¹H-NMR (CDCl₃) δ: 1.32 (9H, s), 2.55 (1H, dd, J=13.3, 6.0 Hz), 2.80 (1H,dd, J=13.3, 3.5 Hz), 3.99 (1H, d, J=10.1 Hz), 4.47 (2H, m), 7.13-7.33(10H, m).

Third Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (488 mg, 1.53mmol) and 43C (500 mg, 1.53 mmol) were added to toluene (8 ml), and themixture was stirred at 110° C. for 1 hour. After the solvent wasdistilled off, the resulting crude product was purified by silica gelcolumn chromatography (chloroform-methanol, 97:3→96:4→94:6, v/v) toobtain 667 mg of Compound 43D as a pale yellow gummy substance.

¹H-NMR (CDCl₃) δ: 1.28 (9H, s), 3.63 (3H, s), 3.80 (1H, m), 3.87 (3H,s), 4.02 (1H, dd, J=14.5, 10.1 Hz), 4.21 (1H, d, J=10.4 Hz), 4.47 (2H,m), 5.20 (1H, d, J=10.8 Hz), 5.26 (1H, d, J=10.7 Hz), 7.30 (15H, m),8.05 (1H, s).

MS: m/z=627 [M+H]⁺.

Fourth Step

To Compound 43D (664 mg, 1.06 mmol) was added 4N HCl (ethyl acetatesolution, 10 ml), and the mixture was stirred at room temperature for 1hour. After the solvent was distilled off under reduced pressure, THFand an aqueous saturated sodium bicarbonate solution were added, and themixture was stirred for 2.5 hours. This was extracted with chloroform,and dried with sodium sulfate. After the solvent was distilled off underreduced pressure, methylene chloride-ethyl ether were added, and theprecipitated solid was filtered off to obtain 458 mg of Compound 43E asa colorless solid.

¹H-NMR (CDCl₃) δ: 3.86 (3H, m), 3.92 (3H, s), 4.41-4.48 (1H, m), 5.32(1H, d, J=10.8 Hz), 5.42 (1H, d, J=10.1 Hz), 5.92 (1H, s), 7.21-7.39(13H, m), 7.59 (2H, m), 7.89 (1H, s).

MS: m/z=495 [M+H]⁺.

Fifth Step

Compound 43E (50 mg, 0.10 mmol) was dissolved in DMF (1 ml), and cesiumcarbonate (165 mg, 0.51 mmol) was added. After stirring at roomtemperature for 30 minutes, iodomethane (0.025 ml, 0.40 mmol) was added,and the mixture was stirred at room temperature for 1 hour. The reactionsolution was poured into water, and this was extracted with ethylacetate, and dried with sodium sulfate. After the solvent was distilledoff under reduced pressure, the resulting crude product was purified bysilica gel column chromatography (chloroform-methanol, 97:3→95:5, v/v)to obtain 60 mg of Compound 43F as a colorless solid.

¹H-NMR (CDCl₃) δ: 2.57 (3H, s), 3.75 (2H, d, J=11.3 Hz), 3.93 (3H, s),4.20-4.29 (2H, m), 5.25 (1H, d, J=9.9 Hz), 5.57 (1H, d, J=9.9 Hz),7.15-7.41 (13H, m), 7.63 (1H, s), 7.72-7.76 (2H, m).

Sixth Step

Compound 43F obtained in Fifth step was dissolved in THF (0.5 ml) andmethanol (0.5 ml), a 2N aqueous sodium hydroxide solution (0.25 ml, 0.50mmol) was added at room temperature, and the mixture was stirred as itwas for 1 hour. 1N hydrochloric acid was added, and this was extractedwith ethyl acetate, and dried with sodium sulfate. After the solvent wasdistilled off under reduced pressure, Compound 43G was obtained as acolorless gummy substance.

Seventh Step

Trifluoroacetic acid (2 ml) was added to Compound 43G obtained in Sixthstep, and the mixture was stirred at room temperature for 1 hour. Afterconcentration under reduced pressure, a pH was adjusted to 3 with anaqueous sodium bicarbonate solution and 2N hydrochloric acid, and thiswas extracted with chloroform, and dried with sodium sulfate. After thesolvent was distilled off under reduced pressure, chloroform-ethyl etherwere added, and the precipitated solid was filtered off to obtain 27 mgof Compound 43 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 2.53 (3H, s), 4.26 (1H, d, J=10.9 Hz), 4.35 (1H, d,J=13.3 Hz), 4.58 (1H, dd, J=13.8, 3.5 Hz), 5.06 (1H, d, J=10.9 Hz), 7.36(10H, m), 8.36 (1H, s), 12.58 (1H, s), 15.62 (1H, s).

MS: m/z=405 [M+H]⁺.

Example 15

First Step

A solution of Compound 49A (2.97 g, 10.4 mmol) in methylene chloride (20ml) was added dropwise to Dess-Martin Periodinane (0.3M, methylenechloride solution, 52.0 ml, 15.6 mmol) at 0° C. After stirring at roomtemperature for 3 hours, this was poured into a 1N aqueous sodiumhydroxide solution, followed by extraction with ethyl ether. The organiclayer was washed with a 1N aqueous sodium hydroxide solution andsaturated sodium chloride water, and dried with magnesium sulfate. Afterthe solvent was distilled off under reduced pressure, 2.08 g of Compound49B was obtained as a white solid.

¹H-NMR (CDCl₃) δ: 3.13 (2H, d, J=6.6 Hz), 4.53 (1H, q, J=6.7 Hz), 5.12(2H, s), 5.28 (1H, brs), 7.26 (10H, m), 9.64 (1H, s).

Second Step

Compound 49B (700 mg, 2.47 mmol), 2-aminoethanol (166 mg, 2.72 mmol) andsodium sulfate (1.76 g, 12.4 mmol) were added to toluene (20 ml), andthe mixture was stirred at room temperature for 1 hour. Boc₂O (0.631 ml,2.72 mmol) was added at room temperature, and the mixture was stirred asit was for 18 hours. The reaction solution was filtered, and thefiltrate was concentrated under reduced pressure. The resulting crudeproduct was purified by silica gel column chromatography (n-hexane-ethylacetate, 1:1, v/v) to obtain 893 mg of 49C as a colorless gummysubstance.

Third Step

Compound 49C (890 mg, 2.09 mmol) and palladium-active carbon (10%, wet,200 mg) were added to ethanol (20 ml), and the mixture was stirred atroom temperature for 2 hours. After filtration with Celite, the solventwas concentrated under reduced pressure to obtain 656 mg of 49D as acolorless oily substance.

¹H-NMR (CDCl₃) δ: 1.40 (9H, s), 2.65-2.86 (2H, m), 3.32 (2H, m), 3.80(2H, m), 4.03-4.12 (1H, m), 4.86 (1H, brs), 7.22 (5H, m).

Fourth Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (610 mg, 2.09mmol) and 49D (664 mg, 2.09 mmol) were added to toluene (6 ml), and themixture was stirred at 100° C. for 4 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (n-hexane-ethyl acetate,1:1, v/v) to obtain 884 mg of Compound 49E as a pale yellow gummysubstance.

MS: m/z=593 [M+H]⁺.

Fifth Step

To Compound 49E (860 mg, 1.45 mmol) was added 4N HCl (ethyl acetatesolution, 10 ml). After stirring at room temperature for 30 minutes, thesolvent was distilled off under reduced pressure. Subsequently, toluene(10 ml) and 2-aminoethanol (0.175 ml, 2.90 mmol) were added, and themixture was stirred at 80° C. for 30 minutes. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol,99:1→95:5→90:10, v/v) to obtain 157 mg of Compound 49F as a colorlessgummy substance and 217 mg of Compound 49G as a yellow solid.

49F: ¹H-NMR (CDCl₃) δ: 2.48 (1H, dd, J=14.0, 11.4 Hz), 3.22 (1H, dd,J=14.1, 3.3 Hz), 3.69 (1H, m), 3.77 (3H, s), 3.83-3.95 (1H, m), 4.08(1H, m), 4.29 (1H, m), 4.41 (1H, m), 5.34 (2H, m), 5.48 (1H, d, J=10.1Hz), 6.86 (2H, m), 7.20-7.39 (7H, m), 7.64 (2H, m)

49G: ¹H-NMR (DMSO-d6) δ: 3.70 (2H, t, J=5.3 Hz), 3.73 (3H, s), 3.86 (2H,t, J=5.3 Hz), 4.14 (2H, s), 4.98 (1H, t, J=5.0 Hz), 5.06 (2H, s), 6.98(1H, s), 7.35 (8H, m), 7.62 (2H, d, J=7.1 Hz), 8.34 (1H, d, J=0.8 Hz).

Sixth Step

Compound 49G (214 mg, 0.465 mmol) was dissolved in THF (4 ml), ethanol(2 ml) and methylene chloride (2 ml), a 2N aqueous sodium hydroxidesolution (1.16 ml, 2.32 mmol) was added at room temperature, and themixture was stirred as it was for 2.5 hours. 1N hydrochloric acid wasadded, and this was extracted with chloroform, and dried with sodiumsulfate. The solvent was distilled off under reduced pressure to obtain158 mg of Compound 49H as a yellow solid.

¹H-NMR (DMSO-d6) δ: 3.70 (2H, q, J=5.2 Hz), 3.89 (2H, t, J=5.3 Hz), 4.22(2H, s), 4.97 (1H, t, J=5.6 Hz), 5.12 (2H, s), 7.23-7.41 (9H, m), 7.60(2H, m), 8.54 (1H, s).

Seventh Step

Compound 49H (50.0 mg, 0.112 mmol) and palladium-active carbon (10%,wet, 12 mg) were added to methanol (1 ml) and DMF (3 ml), and themixture was stirred at room temperature for 5 hours under a hydrogenatmosphere. After filtration with Celite, the solvent was concentratedunder reduced pressure. Chloroform-methanol-ethyl ether were added, andthe precipitated solid was filtered off to obtain 9.0 mg of Compound 49as a colorless solid.

¹H-NMR (DMSO-d6) δ 3.10 (2H, m), 3.51-3.69 (4H, m), 4.10 (1H, d, J=10.7Hz), 4.94 (2H, m), 7.11-7.26 (5H, m), 8.03 (1H, s), 12.94 (1H, brs),15.30 (1H, brs). MS: m/z=359 [M+H]⁺

Example 16

First Step

Compound 50A (1.00 g, 3.98 mmol), triphenylphosphine (1.15 g, 4.48 mmol)and N-methyl-2-nitrobenzenesulfonamide (860 mg, 3.98 mmol) were added toTHF (10 ml), and diethyl azodicarboxylate (2.2M in toluene, 1.99 ml,4.38 mmol) was added dropwise at room temperature. After stirring atroom temperature for 3 hours, the solvent was distilled off underreduced pressure. The resulting crude product was purified by silica gelcolumn chromatography (n-hexane-ethyl acetate, 1:1, v/v) to obtain 710mg of Compound 50B as a colorless gummy substance.

Second Step

Compound 50B (458 mg, 1.02 mmol) was dissolved in acetonitrile,potassium carbonate (422 mg, 3.06 mmol) and benzenethiol (0.126 ml, 1.22mmol) were added, and the mixture was stirred at room temperature for 5hours. The reaction solution was poured into a 1N aqueous sodiumhydroxide solution, and this was extracted with methylene chloride, anddried with sodium sulfate. The resulting crude product was purified byamino column chromatography (chloroform-methanol, 95:5, v/v) to obtain147 mg of Compound 50C as a colorless oily substance.

¹H-NMR (CDCl₃) δ: 1.36 (9H, s), 2.40 (3H, s), 2.51-2.89 (4H, m), 3.90(1H, s), 4.69 (1H, s), 7.17-7.31 (5H, m).

Third Step

Compound 50C (140 mg, 0.530 mmol) and3-(benzyloxy)-4-oxo-4H-pyran-2-carboxylic acid (WO 2006/116764, 119 mg,0.482 mmol) were added to THF (3 ml),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (111 mg,0.578 mmol) and 1-hydroxybenzotriazole (65.1 mg, 0.482 mmol) were added,and the mixture was stirred at room temperature for 18 hours. Thereaction solution was poured into an aqueous sodium bicarbonatesolution, and this was extracted with ethyl acetate, and dried withsodium sulfate. The resulting crude product was purified by silica gelcolumn chromatography (chloroform-methanol, 97:3, v/v) to obtain 219 mgof Compound 50D as a colorless solid.

MS: m/z=493 [M+H]⁺.

Fourth Step

To Compound 50D (216 mg, 0.439 mmol) was added 4N HCl (ethyl acetatesolution, 3 ml). After the mixture was stirred at room temperature for 1hour, the solvent was distilled off under reduced pressure.Subsequently, ethanol (4 ml) and an aqueous saturated sodium carbonatesolution (3 ml) were added, and mixture was stirred at 60° C. for 2hours. Water was added, and this was extracted with ethyl acetate, anddried with sodium sulfate. The resulting crude product was purified byamino column chromatography (chloroform-methanol, 95:5, v/v) to obtain108 mg of Compound 50E as a pale yellow gummy substance.

¹H-NMR (CDCl₃) δ: 3.00 (2H, m), 3.13 (3H, s), 3.18 (1H, m), 3.88 (1H,dd, J=13.5, 3.4 Hz), 4.00-4.07 (1H, m), 5.26 (1H, d, J=10.2 Hz), 5.46(1H, d, J=10.1 Hz), 6.25 (1H, d, J=7.5 Hz), 6.73 (1H, d, J=7.5 Hz),6.99-7.02 (2H, m), 7.28-7.37 (6H, m), 7.63-7.67 (2H, m).

Fifth Step

Trifluoroacetic acid (2 ml) was added to Compound 50E (105 mg, 0.280mmol), and the mixture was stirred at room temperature for 30 minutes.After concentration under reduced pressure, a pH was adjusted to 6 withan aqueous sodium bicarbonate solution and 2N hydrochloric acid, andthis was extracted with chloroform, and dried with sodium sulfate. Afterthe solvent was distilled off under reduced pressure, methylenechloride-methanol-ethyl ether were added, and the precipitated solid wasfiltered off to obtain 29 mg of Compound 50 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 2.99 (3H, s), 3.26-3.47 (3H, m), 4.07 (1H, d, J=11.1Hz), 4.80 (1H, m), 6.43 (1H, d, J=6.9 Hz), 7.11-7.29 (5H, m), 7.50 (1H,d, J=6.9 Hz).

MS: m/z=285 [M+H]⁺.

Example 17

First Step

A solution of Compound 65A (WO 2006/088173, 20.0 g, 69.6 mmol) in THF(1.1 L) was retained at 25° C. on a water bath, and a solution of sodiumchlorite (25.2 g, 278 mmol) and amidosulfuric acid (27.0 g, 278 mmol) inwater (378 mL) was added dropwise over 30 minutes. The reaction solutionwas stirred at the same temperature for 1 hour, and concentrated underreduced pressure. To the residue were added ice water (100 mL) anddiethyl ether (100 mL), and the precipitated solid was filtered. Theresulting crude purified product was washed with water and diethyl etherto obtain 20.3 g of Compound 65B as a white solid.

¹H NMR (DMSO-d6) δ: 3.74 (3H, s), 5.11 (2H, s), 7.31-7.38 (3H, m), 7.48(2H, d, J=7.2 Hz), 8.11 (1H, s), 12.07 (1H, brs).

Second Step

Compound 65B (2.0 g, 6.59 mmol) was dissolved in DMF (340 mL), and HATU(2.76 g, 7.25 mmol), methylamine (2 mol/L THF solution, 3.63 mL, 7.25mmol), and triethylamine (9.89 mmol) were added, and the mixture wasstirred at room temperature for 5 hours. The reaction solution wasdispensed to ethyl acetate and water. The ethyl acetate layer wasseparated, and the aqueous layer was extracted with ethyl acetate once.The combined extracts were washed with water and saturated sodiumchloride water, and dried. The solvent was distilled off to obtain 1.66g of a crude purified product of Compound 65C as a white solid.

¹H-NMR (DMSO-d6) δ: 3.38 (3H, brs), 3.75 (3H, s), 5.37 (2H, s),7.34-7.44 (5H, m), 8.10 (1H, s), 8.38 (1H, s), 11.84 (1H, brs).

Third Step

Potassium carbonate (1.04 g, 7.59 mmol) andO-(2,4-dinitrophenyl)hydroxylamine (831 mg, 4.17 mmol) were added to asolution of Compound 65C (1.2 g, 3.79 mmol) in DMF (20 mL), and themixture was stirred at room temperature for 3 hours. Water was added tothe reaction solution, and the precipitated solid was filtered off andwashed with water to obtain 1.0 g of a crude purified product ofCompound 65D.

¹H-NMR (DMSO-d₆) δ: 3.74 (3H, s), 3.83 (3H, brs), 5.05 (2H, s), 6.46(2H, brs), 7.31-7.38 (5H, m), 8.20 (1H, s), 8.52 (1H, brs).

Fourth Step

Paraformaldehyde (109 mg, 3.62 mmol) and acetic acid (0.017 ml, 0.302mmol) were added to a solution of Compound 65D (1.0 g, 3.02 mmol) in DMF(10 mL) at room temperature, and the mixture was stirred at 105° C. for2 hours. The reaction solution was cooled to 0° C., cesium carbonate(3.44 g, 10.6 mmol) was added, and the mixture was stirred at roomtemperature for 1 hour. Water was added to the reaction solution, andthis was dispensed to ethyl acetate and water. The organic layer waswashed with saturated sodium chloride water, and dried. The solvent wasdistilled off to obtain 120 mg of Compound 65E.

MS: m/z=344 [M+H]⁺.

Fifth Step

Cesium carbonate (81.4 mg, 0.25 mmol) and methylamine (2 mol/L THFsolution, 0.125 ml, 0.25 mmol) were added to a solution of Compound 65E(17.0 mg, 0.05 mmol) in DMF (1 mL), and the mixture was stirred at roomtemperature for 5 hours. The reaction solution was filtered, and thefiltrate was taken and purified by LCMS to obtain Compound 65F.

MS: m/z=358 [M+H]⁺.

Sixth Step

A 2N aqueous sodium hydroxide solution (0.2 mL) was added to a solutionof Compound 65F in DMF (0.5 mL), and the mixture was stirred at roomtemperature for 2 hours Ion-exchange resin DOWEX (50W-X8) was added tothe reaction solution, and this was filtered, and washed with DMF. Afterthe filtrate was concentrated, trifluoroacetic acid (0.5 mL) was added,and the mixture was stirred at 80° C. for 4 hours. After the reactionsolution was concentrated, water and chloroform were added, and theorganic layer was separated. The organic layer was concentrated, andtaken and purified by LCMS to obtain 6.47 mg of Compound 65.

MS: m/z=254 [M+H]⁺.

Example 18

First Step

Compound 95A (WO2006/116764, 1 g, 4.06 mmol) was dissolved in 28%aqueous ammonia, and the solution was stirred at room temperature for 12hours. After the reaction solution was concentrated, the resultingresidue was neutralized with 2N hydrochloric acid, and the precipitatedsolid was suspended in ethyl acetate, filtered off, and dried to obtain1.14 g (yield 100%) of Compound 95B.

¹H-NMR (DMSO-d6) δ: 5.14 (2H, s), 7.31 (1H, d, J=6.6 Hz), 7.34-7.41 (3H,m), 7.45-7.51 (2H, m), 8.17 (1H, d, J=6.6 Hz).

Second Step

WSC HCl (3.06 g, 15.98 mmol) and HOBt (1.58 g, 11.7 mmol) were added toa solution of Compound 95B (3.00 g, 10.65 mmol) in DMF (10 ml) at roomtemperature, the mixture was stirred for 10 minutes, and a methylamine33 wt. % ethanol solution (1.50 g, 15.98 mmol) was added dropwise. Afterthe reaction solution was stirred at the same temperature for 2 hours,and water was added, followed by extraction with chloroform five times.The extract was dried with sodium sulfate, the solvent was distilledoff, and the resulting oil product was purified by subjecting to silicagel chromatography. From a fraction eluting with ethyl acetate-MeOH(6:4, v/v), 2.62 g (yield 95%) of Compound 95C was obtained as a solid.

¹H-NMR (CDCl₃) δ: 2.77 (3H, d, J=4.8 Hz), 5.49 (2H, s), 6.57 (1H, d,J=6.9 Hz), 7.25-7.43 (5H, m), 7.48 (1H, t, J=6.0 Hz), 8.23 (1H, brs),9.77 (1H, brs).

Third Step

Potassium carbonate (4.20 g, 30.42 mmol) was suspended in a solution ofCompound 95C (2.62 g, 10.14 mmol) in DMF (10 ml) at room temperature,the suspension was stirred for 5 minutes,O-(2,4-dinitrophenyl)hydroxylamine (3.03 g, 15.21 mmol) was added, andthe mixture was stirred at the same temperature for 3 hours. Water wasadded to the reaction solution, this was extracted with chloroform fivetimes, and the extract was dried with sodium sulfate. After the solventwas distilled off, the resulting oil product was purified by subjectingto silica gel chromatography. From a faction eluting with ethylacetate-MeOH (6:4, v/v), 1.41 g (yield 51%) of Compound 95D was obtainedas a solid.

¹H-NMR (CDCl₃) δ: 2.62 (3H, d, J=5.1 Hz), 5.06 (2H, s), 5.22 (2H, s),6.18 (1H, d, J=7.8 Hz), 7.25-7.36 (5H, m), 5.89 (1H, d, J=7.8 Hz), 7.57(1H, q, J=5.1 Hz).

Fourth Step

Paraformaldehyde (109.9 mg, 3.66 mmol) and acetic acid (22 mg, 0.37mmol) were added to a solution of Compound 95D (1.0 g, 3.66 mmol) intoluene (10 ml), and the mixture was heated and stirred at 100° C. for40 minutes. After cooling, the solvent was distilled off, the residuewas dissolved in DMF (10 ml) without purification, cesium carbonate(3.58 g, 10.98 mmol) was added under ice cooling, and the mixture wasstirred for 10 minutes. Benzhydryl bromide (1.36 g, 5.49 mmol) was addedto the reaction solution, the mixture was stirred at room temperaturefor 3 hours, and water was added, followed by extraction with ethylacetate three times. The extract was washed with water three times, anddried with sodium sulfate. The solvent was distilled off, and theresulting oil product was purified by subjecting to silica gelchromatography. From a fraction eluting with ethyl acetate-MeOH (9:1,v/v), 1.26 g (yield 71%) of Compound 95E was obtained as a solid.

¹H-NMR (CDCl₃) δ: 2.91 (3H, s), 4.26 (1H, d, J=13.2 Hz), 4.77 (1H, d,J=13.2 Hz), 5.12 (1H, s), 5.42 (1H, J=13.2 Hz), 5.45 (1H, d, J=13.2 Hz),5.82 (1H, J=7.5 Hz), 6.71 (1H, d, J=7.5 Hz), 7.10-7.23 (5H, m),7.27-7.46 (6H, m), 7.52 (2H, d, J=6.9 Hz), 7.60-7.64 (2H, m).

Fifth Step

Compound 95E (100 mg, 0.221 mmol) was dissolved in trifluoroacetic acid(2 ml) and the solution was stirred at room temperature for 1 hour. Thesolvent was distilled off, and the residue was dissolved indichloromethane (2 ml), and neutralized with an aqueous saturated sodiumbicarbonate solution. The resulting solution was made acidic with anaqueous citric acid solution, and the organic layer was separated. Theaqueous layer was extracted with dichloromethane once, and the combinedorganic layers were washed with water, and dried with sodium sulfate.After the solvent was distilled off, the resulting solid was washed withdiisopropyl ether to obtain 50 mg (yield 63%) of Compound 95.

¹H-NMR (CDCl₃) δ: 2.95 (3H, s), 4.36 (1H, d, J=13.2 Hz), 4.95 (1H, d,J=13.2 Hz), 5.22 (1H, s), 5.71 (1H, d, J=7.8 Hz), 6.75 (1H, d, J=7.8Hz), 7.21 (5H, br s), 7.33-7.47 (4H, m), 7.55 (2H, d, J=6.6 Hz).

Example 19

First Step

Paraformaldehyde (299 mg, 9.96 mmol) and acetic acid (1 ml) were addedto a solution of Compound 107A (3.0 g, 9.96 mmol) synthesized accordingto the synthetic method of Compound 95D in DMF (30 ml), and the mixturewas heated and stirred at 120° C. for 4 hours. After the solvent wasdistilled off, ethyl acetate-diisopropyl ether were added to theresidue, and the precipitated solid was filtered off to obtain 2.85 g(yield 91%) of Compound 107B.

¹H-NMR (CDCl₃) δ: 1.19 (6H, J=6.6 Hz), 4.34 (2H, J=7.5 Hz), 4.72-4.86(1H, m), 5.30 (2H, s), 5.49 (1H, t, J=7.5 Hz), 6.36 (1H, d, J=7.8 Hz),7.26-7.35 (4H, m), 7.37 (1H, d, J=7.8 Hz), 7.55-7.58 (2H, m).

Second Step

To a solution of Compound 107B (100 mg, 0.319 mmol) in acetic acid (2ml) were added 96% sulfuric acid (0.5 ml) andbis(3-chlorophenyl)methanol (242.3 mg, 0.957 mmol) at room temperature,and the mixture was stirred at 80° C. for 2 hours. After the reactionsolution was cooled to room temperature, water was added, followed byextraction with ethyl acetate three times. The organic layer was washedwith water once, and dried with sodium sulfate. After the solvent wasdistilled off, diisopropyl ether was added to the residue, and theprecipitated solid was filtered off to obtain 42 mg (yield 29%) ofCompound 107.

¹H-NMR (CDCl₃) δ: 0.953 (3H, d, J=3.9 Hz), 1.12 (3H, d, J=4.2 Hz), 4.51(1H, 13.5 Hz), 4.83 (1H, d, J=13.5 Hz), 4.83-4.92 (1H, m), 5.18 (1H, s),5.74 (1H, d, J=7.8 Hz), 6.73 (1H, d, J=7.8 Hz), 6.90 (1H, d, J=7.5 Hz),7.12 (2H, dd, J=7.2 Hz, 8.1 Hz), 7.19-7.22 (1H, m), 7.37-7.41 (3H, m),7.55 (1H, 5).

Example 20

First Step

Compound 95B (1.00 g, 3.55 mmol) and cyclopropanamine (0.492 ml, 7.10mmol) were added to pyridine (20 ml), 1-hydroxybenzotriazole (544 mg,3.55 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (1.36 g, 7.10 mmol) were sequentially added, and themixture was stirred at room temperature for 18 hours. The solvent wasdistilled off under reduced pressure, and the resulting crude productwas purified by silica gel column chromatography (chloroform methanol,95:5, v/v) and, subsequently, amino column chromatography(chloroform-methanol, 99:1, v/v) to obtain 1.19 g of Compound 126A as acolorless solid.

¹H-NMR (CDCl₃) δ: 0.22 (1H, m), 0.70 (2H, m), 2.76-2.83 (1H, m), 5.50(2H, s), 6.59 (1H, dd, J=7.0, 1.9 Hz), 7.44 (5H, d, J=0.7 Hz), 7.53 (1H,dd, J=6.9, 6.2 Hz), 8.30 (1H, brs), 9.71 (1H, brs).

Second Step

Compound 126A (1.19 g, 4.19 mmol) was dissolved in DMF (15 ml),potassium carbonate (2.90 g, 20.1 mmol) was added, and the mixture wasstirred at room temperature for 30 minutes.O-(2,4-dinitrophenyl)hydroxylamine (1.67 g, 8.38 mmol) was added, andthe mixture was stirred at room temperature for 18 hours. Chloroform wasadded to the reaction solution, the precipitated yellow precipitate wasfiltered to remove, and the filtrate was concentrated under reducedpressure. The resulting crude product was purified by amino columnchromatography (chloroform-methanol, 97:3→95:5, v/v) to obtain 851 mg ofCompound 126B as a yellow solid.

¹H-NMR (CDCl₃) δ: 0.41-0.46 (2H, m), 0.76 (2H, m), 2.73-2.81 (1H, m),5.19 (2H, s), 5.61 (2H, s), 6.26 (1H, d, J=7.2 Hz), 7.38 (5H, s), 7.44(1H, d, J=7.8 Hz), 7.70 (1H, s).

Third Step

Compound 126B (847 mg, 2.83 mmol) and paraformaldehyde (255 mg, 8.49mmol) were added to ethanol (12 ml), and the mixture was stirred at 140°C. for 30 minutes under microwave irradiation, the resulting solutionwas concentrated under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol,97:3→95:5→90:10, v/v) and, subsequently, amino column chromatography(chloroform-methanol, 97:3, v/v) methylene chloride-ethyl ether wereadded, and the precipitated solid was filtered off to obtain 665 mg ofCompound 126C as a colorless solid.

¹H-NMR (CDCl₃) δ: 0.61-0.66 (2H, m), 0.87 (2H, m), 2.68-2.76 (1H, m),4.32 (2H, d, J=7.9 Hz), 5.28 (2H, s), 6.33 (1H, d, J=7.7 Hz), 6.45 (1H,t, J=7.7 Hz), 7.33 (3H, m), 7.38 (1H, d, J=7.7 Hz), 7.52 (2H, m).

Fourth Step

Compound 126C (100 mg, 0.321 mmol) was dissolved in DMF (0.5 ml), cesiumcarbonate (314 mg, 0.964 mmol) and (bromomethylene)dibenzene (119 mg,0.482 mmol) were added at 0° C., and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was poured into water,this was extracted with ethyl acetate, and the organic layer was washedwith water, and dried with sodium sulfate. The solvent was distilled offunder reduced pressure, and the resulting crude product was purified bysilica gel column chromatography (chloroform-methanol, 97:3→95:5, v/v)to obtain 124 mg of Compound 126D as a colorless gummy substance.

¹H-NMR (CDCl₃) δ: 0.37-0.47 (2H, m), 0.74 (2H, m), 2.63-2.68 (1H, m),4.35 (1H, d, J=13.4 Hz), 4.65 (1H, d, J=13.4 Hz), 5.07 (1H, s), 5.40(1H, d, J=10.7 Hz), 5.47 (1H, d, J=10.5 Hz), 5.79 (1H, d, J=7.6 Hz),6.67 (1H, d, J=7.8 Hz), 7.04-7.62 (15H, m).

Fifth Step

Trifluoroacetic acid (2 ml) was added to Compound 126D obtained inFourth step, and the mixture was stirred at room temperature for 1.5hours. After concentrated under reduced pressure, a pH was adjusted to 6with an aqueous sodium bicarbonate solution and 2N hydrochloric acid,and this was extracted with chloroform and dried with sodium sulfate.After the solvent was distilled off under reduced pressure, methylenechloride-ethyl ether were added, and the precipitated solid was filteredoff to obtain 52 mg of Compound 126 as a colorless solid.

¹H-NMR (DMSO-d₆) δ: −0.19-0.06 (1H, m), 0.44-0.54 (1H, m), 0.82 (2H, m),2.62-2.69 (1H, m), 4.21 (1H, d, J=13.3 Hz), 5.11 (1H, d, J=13.1 Hz),5.32 (1H, s), 5.47 (1H, t, J=11.1 Hz), 7.13 (1H, d, J=7.6 Hz), 7.23 (3H,m), 7.28-7.47 (8H, m), 7.69 (2H, t, J=8.5 Hz).

MS: m/z=388 [M+H]⁺.

Example 21

First Step

Compound 95B (2.40 g, 8.52 mmol) and ethyl 3-aminopropanoatehydrochloride (2.62 g, 17.0 mmol) were added to pyridine (30 ml),1-hydroxybenzotriazole (1.31 g, 8.52 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.27 g,17.0 mmol) were sequentially added, and the mixture was stirred at roomtemperature for 2 hours. The solvent was distilled off under reducedpressure, and the resulting crude product was purified by amino columnchromatography (chloroform-methanol, 95:5, v/v) to obtain 1.90 g ofCompound 128A as a colorless solid.

¹H-NMR (CDCl₃) δ: 1.29 (3H, t, J=7.1 Hz), 2.48 (2H, t, J=6.4 Hz), 3.58(2H, q, J=6.3 Hz), 4.17 (2H, q, J=7.1 Hz), 5.59 (2H, s), 6.57 (1H, dd,J=7.1, 1.6 Hz), 7.37-7.52 (6H, m), 8.73 (1H, brs), 9.72 (1H, brs).

Second Step

Compound 128A (2.58 g, 7.49 mmol) was dissolved in DMF (30 ml),potassium carbonate (5.18 g, 37.5 mmol) was added, and the mixture wasstirred at room temperature for 30 minutes.O-(2,4-dinitrophenyl)hydroxylamine (2.98 g, 15.0 mmol) was added, andthe mixture was stirred at room temperature for 20 hours. Chloroform wasadded to the reaction solution, the precipitated yellow precipitate wasfiltered to remove, and the filtrate was concentrated under reducedpressure. The resulting crude product was purified by amino columnchromatography (chloroform-methanol, 97:3→95:5, v/v) and, subsequently,silica gel column chromatography (chloroform-methanol, 95:5→92:8, v/v)to obtain 1.67 g of Compound 128B as a yellow solid.

¹H-NMR (CDCl₃) δ: 1.26 (3H, t, J=7.2 Hz), 2.42 (2H, t, J=6.6 Hz), 3.43(2H, q, J=6.4 Hz), 4.12 (2H, q, J=7.1 Hz), 5.13 (2H, s), 5.53 (2H, s),6.21 (1H, d, J=7.6 Hz), 7.33 (5H, s), 7.39 (1H, d, J=7.6 Hz), 7.85 (1H,t, J=5.6 Hz).

Third Step

Compound 128B (1.66 g, 4.62 mmol) and paraformaldehyde (416 mg, 13.9mmol) were added to ethanol (20 ml), and the mixture was stirred at 140°C. for 30 minutes under microwave irradiation. The reaction solution wasconcentrated under reduced pressure, and the resulting crude product waspurified by amino column chromatography (chloroform-methanol,99:1→3.95:5, v/v) to obtain 1.57 g of Compound 128C as a colorlesssolid.

¹H-NMR (CDCl₃) δ: 1.27 (3H, t, J=7.2 Hz), 2.70 (2H, t, J=5.7 Hz), 3.57(2H, t, J=5.8 Hz), 4.13 (2H, q, J=7.1 Hz), 4.50 (2H, d, J=7.9 Hz), 5.27(2H, s), 5.87 (1H, t, J=7.8 Hz), 6.32 (1H, d, J=7.6 Hz), 7.31 (4H, m),7.54 (2H, m).

Fourth Step

Compound 128C (1.00 g, 2.69 mmol) was dissolved in DMF (10 ml), cesiumcarbonate (2.63 g, 8.08 mmol) and (bromomethylene)dibenzene (998 mg,4.04 mmol) were added at 0° C., and the mixture was stirred at roomtemperature for 18 hours. The reaction solution was poured into water,this was extracted with ethyl acetate, and the organic layer was washedwith water, and dried with sodium sulfate. The solvent was distilled offunder reduced pressure, and the resulting crude product was purified bysilica gel column chromatography (chloroform-methanol, 98:2, v/v) toobtain 500 mg of Compound 128D as a colorless gummy substance.

¹H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.3 Hz), 2.46 (1H, m), 2.70-2.80 (1H,m), 2.87-2.96 (1H, m), 4.11 (2H, q, J=7.3 Hz), 4.12 (1H, m), 4.48 (1H,d, J=13.7 Hz), 4.85 (1H, d, J=13.7 Hz), 5.10 (1H, s), 5.47 (2H, s), 5.83(1H, d, J=8.0 Hz), 6.73 (1H, d, J=8.0 Hz), 7.37 (15H, m).

Fifth Step

Triflouroacetic acid (1 ml) was added to Compound 128D (40 mg, 0.074mmol), and the mixture was stirred at room temperature for 1 hour. Afterconcentrated under reduced pressure, a pH was adjusted to 6 with anaqueous sodium bicarbonate solution and 2 N hydrochloric acid, and thiswas extracted with chloroform, and dried with sodium sulfate. After thesolvent was distilled off under reduced pressure, methylenechloride-ethyl ether were added, and the precipitated solid was filteredoff to obtain 20 mg of Compound 128 as a colorless solid.

¹H-NMR (DMSO-d₆) δ: 1.16 (3H, t, J=7.1 Hz), 2.45-2.58 (3H, m), 3.70 (1H,m), 4.02 (2H, q, J=7.1 Hz), 4.39 (1H, d, J=13.4 Hz), 5.09 (1H, d, J=13.3Hz), 5.48 (1H, d, J=3.2 Hz), 5.51 (1H, s), 7.19-7.38 (7H, m), 7.45 (2H,t, J=7.3 Hz), 7.69 (2H, d, J=7.2 Hz).

MS: m/z=448 [M+H]⁺.

Example 22

First Step

Compound 95B (1.50 g, 5.32 mmol) and tert-butyl2-aminoethyl(methyl)carbamate (1.86 g, 10.7 mmol) were added to pyridine(20 ml), 1-hydroxybenzotriazole (815 mg, 5.32 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.04 g,10.7 mmol) were sequentially added, and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was poured into 1Nhydrochloric acid, this was extracted with ethyl acetate, and dried withsodium sulfate. The solvent was distilled off under reduced pressure,and the resulting crude product was purified by amino columnchromatography (chloroform-methanol, 95:5, v/v) and, subsequently,silica gel column chromatography (chloroform-methanol, 95:5, v/v) toobtain 1.63 g of Compound 135A as a colorless gummy substance.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 2.82 (3H, s), 3.28 (4H, m), 5.59 (2H,s), 6.57 (1H, d, J=6.0 Hz), 7.46 (6H, m), 8.46 (1H, m), 9.68 (1H, brs).

Second Step

Compound 135A (1.05 g, 2.62 mmol) was dissolved in DMF (15 ml),potassium carbonate (1.81 g, 13.1 mmol) was added, and the mixture wasstirred at room temperature for 30 minutes.O-(2,4-dinitrophenyl)hydroxylamine (1.04 g, 5.23 mmol) was added, andthe mixture was stirred at room temperature for 18 hours. Chloroform wasadded to the reaction solution, the precipitated yellow precipitate wasfiltered to remove, and the filtrate was concentrated under reducedpressure. The resulting crude product was purified by amino columnchromatography (chloroform methanol, 97:3→95:5, v/v) to obtain 887 mg ofCompound 135B as a pale yellow solid.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 2.84 (3H, s), 3.38 (4H, m), 5.33 (2H,s), 5.68 (1H, brs), 5.80 (1H, brs), 6.35 (1H, d, J=7.6 Hz), 6.74 (1H,brs), 7.39 (5H, brm), 7.52 (1H, t, J=9.5 Hz).

Third Step

Compound 135B (880 mg, 2.11 mmol) and paraformaldehyde (190 mg, 6.34mmol) were added to ethanol (18 ml), and the mixture was stirred at 140°C. for 30 minutes under microwave irradiation. The reaction solution wasconcentrated under reduced pressure, and the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol,97:3→95:5→90:10, v/v) and, subsequently, amino column chromatography(chloroform-methanol, 97:3, v/v) to obtain 721 mg of Compound 135C as acolorless solid.

¹H-NMR (CDCl₃) δ: 1.29 (9H, s), 2.95 (3H, s), 4.38 (2H, brs), 5.33 (2H,brs), 6.36 (1H, d, J=7.6 Hz), 6.85 (1H, t, J=7.4 Hz), 7.33 (4H, m), 7.55(2H, m).

MS: m/z=429 [M+H]⁺.

Fourth Step

Compound 135C (720 mg, 1.68 mmol) was dissolved in DMF (3.5 ml), cesiumcarbonate (1.64 g, 5.04 mmol) and (bromomethylene)dibenzene (623 mg,2.52 mmol) were added at 0° C., and the mixture was stirred at roomtemperature for 18 hours. The reaction solution was poured into water,this was extracted with ethyl acetate, and the organic layer was washedwith water, and dried with sodium sulfate. The solvent was distilled offunder reduced pressure, and the resulting crude product was purified bysilica gel column chromatography (chloroform-methanol, 97:3→95:5, v/v)to obtain 732 mg of Compound 135D.

Fifth Step

To Compound 135D (727 mg, 1.22 mmol) was added 4N HCl (ethyl acetatesolution, 10 ml). After stirring at room temperature for 1 hour, thesolvent was distilled off under reduced pressure. An aqueous saturatedsodium bicarbonate solution was added, and this was extracted withchloroform, and dried with sodium sulfate. The solvent was distilled offunder reduced pressure, methylene chloride-ethyl ether were added to theresulting crude product, and the precipitated solid was filtered off toobtain 575 mg of Compound 135E as a colorless solid.

Sixth Step

Trifluoroacetic acid (2 ml) was added to Compound 135E (50 mg, 0.10mmol) and the mixture was stirred at room temperature for 1.5 hours.After concentrated under reduced pressure, a pH was adjusted to 6 withan aqueous sodium bicarbonate solution and an aqueous ammonium chloridesolution, and this was extracted with chloroform, and dried with sodiumsulfate. After the solvent was distilled off under reduced pressure,methylene chloride-ethyl ether were added, and the precipitated solidwas filtered to obtain 15 mg of Compound 135 as a colorless solid.

¹H-NMR (DMSO-d₆) δ: 2.40 (3H, s), 2.80 (1H, s), 3.12 (3H, m), 3.87 (1H,m), 4.37 (1H, d, J=13.6 Hz), 5.10 (1H, d, J=13.4 Hz), 5.52 (1H, s), 5.53(1H, d, J=5.5 Hz), 7.15-7.70 (11H, m).

MS: m/z=405 [M+H]⁺.

Example 23

First Step

Potassium carbonate (17.9 g, 129 mmol) and methyl iodide (8.03 mL, 129mmol) were sequentially added to a solution of Compound 165A (WO2006/088173, 37.0 g, 108 mmol) in DMF (370 mL) at room temperature, andthe mixture was stirred for 1.5 hours. The reaction solution was addedto a solution of ammonium chloride (20.8 g, 390 mmol) in water (1110 mL)under ice-cooling, and the precipitated solid was filtered off, andwashed with water to obtain a crude product (33 g). The aqueous layerwas salted out with sodium chloride, extracted with ethyl acetate, anddried with sodium sulfate. The solvent was distilled off under reducedpressure, and a crude product (9 g) was obtained from the resultingresidue. The crude products were combined, and purified by silica gelcolumn chromatography (ethyl acetate/n-hexane=50%→100%) to obtainCompound 165B (36.5 g, 95%) as a white solid.

Second Step

Potassium osmate dihydrate (1.13 g, 3.06 mmol), sodium periodate (87.3g, 408 mmol) and water (365 mmol) were sequentially added to a solutionof Compound 165B (36.5 g, 102 mmol) in 1,4-dioxane (548 mL) at roomtemperature, and the mixture was stirred for 6 hours. The reactionsolution was extracted with methylene chloride, and dried with sodiumsulfate. The solvent was distilled off under reduced pressure, and theresulting residue was purified by silica gel column chromatography(ethyl acetate/n-hexane=50%→100%) to obtain Compound 165C (33.0 g, 90%)as a brown foam.

Third Step

Ethylenediamine (0.247 mL, 3.66 mmol) and acetic acid (0.0210 mL, 0.366mmol) were sequentially added to a suspension of Compound 165C (1.38 g,3.66 mmol) in toluene (25 mL) at room temperature, and the mixture wasstirred for 1 hour, and further stirred at 50° C. for 17 hours. Theprecipitated solid was filtered off, and washed with ether to obtainCompound 165D (1.11 g, 100%) as a pale yellow solid.

¹HNMR (DMSO-d₆) δ: 3.05 (2H, m), 3.26 (1H, m), 3.63 (2H, m), 3.75 (3H,s), 3.87 (1H, m), 4.52 (1H, dd, J=3.3, 12.6 Hz), 4.69 (1H, m), 4.99 (1H,d, J=10.4 Hz), 5.15 (1H, d, J=10.4 Hz), 7.35 (3H, m), 7.54 (2H, m), 8.41(1H, s).

Fourth Step

Bromodiphenylmethane (2.26 g, 9.14 mmol) was added to a suspension ofCompound 165D (2.77 g, 7.50 mmol), potassium carbonate (2.23 g, 16.1mmol) and sodium iodide (102 mg, 0.680 mmol) in acetonitrile (30 mL) atroom temperature, and the mixture was stirred at 90° C. for 7 hours. Thereaction solution was poured into hydrochloric acid (2 N, 10 mL) and ice(20 g), and this was extracted with chloroform (100 mL×2), and driedwith sodium sulfate. The solvent was distilled off under reducedpressure, and the resulting residue was purified by silica gel columnchromatography (chloroform/methanol=0%→5%) to obtain Compound 165E (2.72g, 68%) as a pale yellow solid.

Fifth Step

An aqueous sodium hydroxide solution (2 N, 10 mL) was added to asolution of Compound 165E (2.72 g, 5.08 mmol) in ethanol (30 mL) at roomtemperature, and the mixture was stirred for 3 days. Hydrochloric acid(1 N, 20 mL) was added to the reaction solution at room temperature(pH=1), and this was extracted with chloroform (100 mL×2), and driedwith sodium sulfate. The solvent was distilled off under reducedpressure, and the resulting residue was purified by silica gel columnchromatography (chloroform/methanol=0%→10%) to obtain Compound 165F(1.77 g, 67%) as a pale yellow solid.

¹HNMR (DMSO-d₆) δ: 2.63 (1H, m), 3.16 (1H, m), 3.49 (1H, m), 3.73 (1H,m), 4.12 (2H, m), 4.56 (1H, m), 5.04 (1H, s), 5.09 (1H, d, J=10.7 Hz),5.19 (1H, d, J=10.7 Hz), 7.28-7.53 (15H, m), 8.32 (1H, s), 8.39 (1H, s).

Sixth Step

A solution of Compound 165F (1.77 g, 3.39 mmol) and lithium chloride(0.515 g, 12.2 mmol) in N,N′-dimethylimidazolidinone (20 mL) was stirredat 90° C. for 1 hour. Water (10 mL), hydrochloric acid (2 N, 10 mL) andwater (10 mL) were sequentially added to the reaction solution at roomtemperature. The precipitated solid was filtered off, and washed withether, DMF-water were added, and the precipitated solid was filtered offto obtain Compound 165 (599 mg, 41%) as a white solid.

¹HNMR (DMSO-d₆) δ: 2.60 (1H, m), 3.20 (1H, m), 3.64 (2H, m), 4.00 (2H,m), 4.55 (1H, m), 5.01 (1H, s), 7.28-7.47 (10H, m), 8.16 (1H, s), 11.97(1H, brs).

MS: m/z=432 [M+H]⁺.

Example 24

First Step

(S)—N1-benzyl-3-phenylpropan-1,2-diamine (Journal of the AmericanChemical Society; English; 127; 30; 2005; 10504, 1.83 g, 7.61 mmol) andacetic acid (0.5 ml) were added to a solution of Compound 167A (WO2006/11674, 3.58 g, 7.61 mmol) in xylene (30 ml), and the mixture wasrefluxed for 2 hours. After cooling to room temperature, the solvent wasdistilled off, and the resulting oil product was purified by subjectingto silica gel chromatography. The column was eluted initially withn-hexane-ethyl acetate (9:1, v/v) and, then, n-hexane-ethyl acetate(1:1, v/v). The objective fraction was concentrated to obtain 349 mg(yield 7%) of Compound 167B.

¹HNMR (CDCl₃) δ: 2.54 (1H, t, J=9.6 Hz), 2.77 (1H, dd, J=9.0 Hz, 13.2Hz), 3.31 (1H, dd, J=6.9 Hz, 9.6 Hz), 3.43-3.78 (5H, m), 4.04-4.15 (1H,m), 4.42-4.48 (1H, m), 4.62 (2H, d, J=6.0 Hz), 5.29 (1H, d, J=10.5 Hz),5.43 (1H, d, J=10.5 Hz), 6.77-6.85 (2H, m), 7.19-7.39 (14H, m), 7.60(2H, d, J=6.3 Hz), 8.05 (1H, s).

Second Step

Boc₂O (3 ml) and DMAP (180 mg, 1.47 mmol) were added to a solution ofCompound 167B (968 mg, 1.47 mmol) in MeCN (10 ml), and the mixture washeated to reflux for 5 hours. A 2N aqueous sodium hydroxide solution wasadded to the reaction solution to stop the reaction, and this wasneutralized using 2N hydrochloric acid, followed by extraction withethyl acetate three times. After the extract was washed with an aqueoussaturated sodium chloride solution, the solvent was distilled off, andthe resulting oil product was purified by silica gel chromatography. Thecolumn was eluted initially with n-hexane-ethyl acetate (6:4, v/v) and,then only ethyl acetate. The objective fraction was concentrated toobtain 349 mg (yield 45%) of 167C as a solid.

¹HNMR (CDCl₃) δ: 2.54 (1H, t=9.0 Hz), 2.76 (1H, dd, J=9.3 Hz, 16.5 Hz),3.31 (1H, dd, J=6.9 Hz, 9.6 Hz), 3.45 (1H, dd, J=3.3 Hz, 12.6 Hz),3.51-3.78 (4H, m), 4.04-4.13 (1H, m), 4.42-4.52 (1H, m), 4.61 (2H, d,J=6.0 Hz), 2.79 (1H, d, J=10.2 Hz), 5.29 (1H, d, J=10.2 Hz), 5.43 (1H,d, J=10.2 Hz), 6.76-7.39 (11H, m), 7.60 (2H, d, J=6.6 Hz), 8.05 (1H, s),10.42 (1H, t, J=5.7 Hz).

Third Step

Compound 167C (150 mg, 0.280 mmol) was dissolved in trifluoroacetic acid(2 ml), and the solution was stirred at room temperature for 1 hour. Thesolvent was distilled off, the residue was dissolved in dichloromethane(2 ml), and the solution was neutralized with an aqueous saturatedsodium bicarbonate solution. The resulting solution was made acidic withan aqueous citric acid solution, and the organic layer was separated.The aqueous layer was extracted with dichloromethane once, and thecombined organic layers were washed with water, and dried with sodiumsulfate. The solvent was distilled off, and the resulting solid waswashed with diisopropyl ether to obtain 71 mg (yield 57%) of Compound167.

¹HNMR (CDCl₃) δ: 2.65 (1H, dd, J=8.4 Hz, 9.6 Hz), 2.97 (1H, dd, J=9 Hz,13.5 Hz), 3.43 (1J, dd, J=7.2 Hz, 9.6 Hz), 3.55 (1H, dd, J=3.0 Hz, 13.2Hz), 3.61-3.80 (4H, m), 4.15 (1H, dd, J=4.2 Hz, 9.9 Hz), 4.51-4.60 (1H,m), 7.15-7.18 (2H, m), 7.28-7.38 (8H, m), 8.02 (1H, s), 12.04 (1H, s).

Example 25

First Step

2,2-Dimethoxyethanamine (0.49 ml, 4.47 mmol) was added to a solution ofCompound 95A (WO 2006/116764, 500 mg, 2.03 mmol) in ethanol (5 mL), andthe mixture was stirred at 80° C. for 3 hours. After the reactionsolution was allowed to cool, acetic acid (0.27 ml, 4.69 mmol) was addedat room temperature, followed by concentration under reduced pressure.The resulting residue was dissolved in DMF (5 mL), and DBU (0.66 mL, 4.4mmol) and, subsequently, methyl iodide (1.02 mL, 16.2 mmol) were addedunder a nitrogen atmosphere, and the mixture was stirred at roomtemperature for 3 hours. An aqueous saturated sodium bicarbonatesolution, and ethyl acetate were added to the reaction solution, theethyl acetate layer was separated, and the aqueous layer was extractedwith ethyl acetate. Sodium sulfate was added to the combined extracts,this was filtered, and concentrated, and the resulting residue waspurified by subjecting to silica gel chromatography. The column waseluted with chloroform-methanol (9:1), and the objective fraction wasconcentrated to obtain 258 mg of Compound 169A as a brown oil.

¹H-NMR (CDCl₃) δ: 3.37 (6H, s), 3.80 (3H, s), 3.87 (2H, d, J=4.8 Hz),4.46 (1H, t, J=4.8 Hz), 5.30 (2H, s), 6.75 (1H, d, J=6.0 Hz), 7.30-7.41(6H, m).

Second Step

Formic acid (31 mL) and, subsequently, water (5 mL) were added toCompound 169A (1.00 g, 2.88 mmol) and the mixture was stirred at 70° C.for 6.5 hours. Water and ethyl acetate were added to the reactionmixture, the ethyl acetate layer was separated, and the aqueous layerwas extracted with ethyl acetate. The combined extracts were washed withan aqueous saturated sodium bicarbonate solution, sodium sulfate wasadded, this was filtered, and concentrated, and the resulting residuewas purified by subjecting to silica gel chromatography. The columnchromatography was eluted with ethyl acetate-methanol, and the objectivefraction was concentrated to obtain a mixture of aldehyde hydrate andmethylacetal as a colorless transparent oil product. The resulting oilproduct was dissolved in dichloromethane (5 mL), 1,3-diaminopropanedihydrochloride (354 mg, 2.41 mmol) and, subsequently, acetic acid(0.069 ml, 1.2 mmol) were added, and the mixture was stirred at roomtemperature for 6 hours. The reaction solution was diluted withdichloromethane, and the insolubles were filtered, and concentratedunder reduced pressure to obtain a crude purified product of Compound169B.

MS: m/z=326.20 [M+H]⁺.

Third Step

Potassium carbonate (498 mg, 3.61 mmol) and, subsequently,bromomethylenedibenzene (890 mg, 3.61 mmol) were added to a solution ofCompound 169B (391 mg, 1.20 mmol) in acetonitrile (4 mL). After thereaction solution was stirred at 90° C. for 2 hours, water, ethylacetate and brine were added to the reaction solution, the ethyl acetatelayer was separated, and the aqueous layer was extracted with ethylacetate once. The combined extracts were dried with magnesium sulfate,filtered, and concentrated. The resulting residue was purified bysubjecting to silica gel column chromatography. The column was elutedwith ethyl acetate-methanol, and the objective fraction was concentratedto obtain 106 mg of Compound 169C as an orange solid.

MS: m/z=492.15 [M+H]⁺.

Fourth Step

Lithium chloride (27.2 mg, 0.641 mmol) was added to a solution ofCompound 169C (105 mg, 0.214 mmol) in DMI (2 mL), and the mixture wasstirred at 90° C. for 3 hours. Lithium chloride (27.2 mg, 0.641 mmol)was further added, and the mixture was stirred at 90° C. for 1 hour. Thereaction solution was concentrated under reduced pressure, and theresulting residue was purified using a LCMS collection apparatus. Theeluted solvent was distilled off, diethyl ether was added to theresidue, and the precipitated solid was filtered off. Washing and dryingwith diethyl ether afforded 27 mg of Compound 169.

¹H-NMR (CD₃OD) δ: 1.63 (1H, dd, J=13.4, 2.8 Hz), 1.84 (1H, br s),2.55-2.64 (1H, m), 2.90-3.10 (2H, m), 4.30 (1H, dd, J=14.5, 4.0 Hz),4.52 (4H, dd, J=14.5, 3.8 Hz), 4.63-4.75 (4H, m), 5.16 (1H, s), 6.16(1H, d, J=7.2 Hz), 6.78 (1H, d, J=7.2 Hz), 7.16-7.32 (10H, m).

MS: m/z=402.10 [M+H]⁺.

Example 26

First Step

Compound 49B (950 mg, 3.35 mmol), 3-aminopropan-1-ol (277 mg, 3.69 mmol)and sodium sulfate (1.91 g, 13.4 mmol) were added to toluene (25 ml),and the mixture was stirred at room temperature for 1 hour. Boc₂O (0.856ml, 3.69 mmol) was added at room temperature, and the mixture wasstirred as it was for 18 hours. Boc₂O (0.400 ml, 1.72 mmol) was furtheradded at room temperature, and the mixture was stirred as it was for 60hours. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure. The resulting crude product waspurified by silica gel column chromatography (n-hexane-ethyl acetate,1:1, v/v) to obtain 1.02 g of 171A as a colorless gummy substance.

Second Step

Compound 171A (1.01 g, 2.29 mmol) and palladium-active carbon (10%, wet,200 mg) were added to ethanol (20 ml), and the mixture was stirred atroom temperature for 1.5 hours under a hydrogen atmosphere. Afterfiltration with Celite, the solvent was concentrated under reducedpressure to obtain 755 mg of 171B as a colorless oily substance.

¹H-NMR (CDCl₃) δ: 1.42 (5H, s), 1.49 (4H, s), 1.56-1.92 (2H, m), 2.49(0.4H, dd, J=13.6, 9.8 Hz), 2.62 (0.6H, dd, J=13.6, 8.5 Hz), 2.81 (0.4H,dd, J=13.5, 3.6 Hz), 3.16 (1.6H, m), 3.60-4.14 (4H, m), 5.13 (0.6H, d,J=8.8 Hz), 5.19 (0.4H, d, J=8.5 Hz), 7.22-7.37 (5H, m).

Third Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (660 mg, 1.99mmol) and 171B (609 mg, 1.99 mmol) were added to toluene (8 ml), and themixture was stirred at 100° C. for 1.5 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 99:1,v/v) to obtain 1.02 g of Compound 171C as a pale yellow gummy substance.

Fourth Step

To Compound 171C (991 mg, 1.60 mmol) was added 4N HCl (ethyl acetatesolution, 12 ml). After the mixture was stirred at room temperature for1 hour, the solvent was distilled off under reduced pressure.Subsequently, toluene (12 ml) and 3-aminopropan-1-ol (0.244 ml, 3.19mmol) were added, and the mixture was stirred at 80° C. for 10 minutes.After the solvent was distilled off under reduced pressure, theresulting crude product was purified by silica gel column chromatography(chloroform-methanol, 99:1→95:5→90:10, v/v) to obtain 341 mg of Compound171D as a yellow gummy substance and 338 mg of Compound 171E as acolorless solid.

171D: ¹H-NMR (CDCl₃) δ: 1.29 (3H, t, J=7.1 Hz), 1.51 (1H, d, J=13.7 Hz),1.97 (1H, m), 2.91 (1H, dd, J=13.8, 9.8 Hz), 2.99-3.10 (2H, m), 3.90(1H, td, J=12.1, 2.5 Hz), 4.12 (2H, m), 4.25 (2H, m), 4.83 (2H, m), 5.33(1H, d, J=10.1 Hz), 5.51 (1H, d, J=10.1 Hz), 6.88 (2H, m), 7.23-7.40(7H, m), 7.68 (2H, m)

171E: ¹H-NMR (CDCl₃) δ: 1.19 (3H, t, J=7.2 Hz), 1.82-1.99 (2H, m), 2.73(1H, dd, J=14.0, 11.3 Hz), 3.13 (1H, m), 3.35 (1H, dd, J=14.0, 3.4 Hz),3.63 (1H, m), 3.90-4.26 (4H, m), 4.43 (1H, d, J=13.6 Hz), 5.27 (1H, t,J=3.5 Hz), 5.31 (2H, s), 6.78 (2H, dd, J=6.3, 3.2 Hz), 7.01 (1H, d,J=7.0 Hz), 7.18 (3H, t, J=3.1 Hz), 7.28-7.39 (3H, m), 7.67 (2H, m).

Fifth Step

Compound 171D (329 mg, 0.673 mmol) was dissolved in ethanol (2 ml) andTHF (4 ml), a 2N aqueous sodium hydroxide solution (1.69 ml, 3.38 mmol)was added, the mixture was stirred at room temperature for 1 hour. Tothe reaction solution was added 2N hydrochloric acid, and this wasextracted with ethyl acetate, and dried with sodium sulfate. The solventwas concentrated under reduced pressure to obtain 215 mg of Compound171F as a colorless solid.

MS: m/z=461 [M+H]⁺.

Sixth Step

Trifluoroacetic acid (2 ml) was added to Compound 171F (50 mg, 0.11mmol), and the mixture was stirred at room temperature for 1 hour. Afterconcentration under reduced pressure, a pH was adjusted to 6 with anaqueous sodium bicarbonate solution and 2N hydrochloric acid, and thiswas extracted with chloroform, and dried with sodium sulfate. After thesolvent was distilled off under reduced pressure,chloroform-methanol-ethyl ether were added, and the precipitated solidwas filtered off to obtain 24 mg of Compound 171 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 1.63 (1H, d, J=12.6 Hz), 1.83 (1H, m), 2.96-3.29(3H, m), 4.05 (2H, m), 4.55 (1H, dd, J=13.2, 4.4 Hz), 5.08 (1H, dd,J=9.2, 5.4 Hz), 5.30 (1H, s), 7.19 (5H, m), 8.09 (1H, s), 12.84 (1H,brs).

MS: m/z=371 [M+H]⁺.

Example 27

First Step

A solution of Compound 2b (1.98 g, 5.48 mmol) in methylene chloride (10ml) was added dropwise to Dess-Martin Periodinane (0.3M, methylenechloride solution, 25.0 ml, 7.50 mmol) at 0° C. After stirring at roomtemperature for 3 hours, the mixture was poured into a 1N aqueous sodiumhydroxide solution, and this was extracted with ethyl ether. The organiclayer was washed with 1N aqueous sodium hydroxide solution and saturatedsodium chloride water, and dried with magnesium sulfate. After thesolvent was distilled off under reduced pressure, this was purified bysilica gel column chromatography (n-hexane-ethyl acetate, 2:1, v/v) toobtain 1.73 g of Compound 175A.

¹H-NMR (CDCl₃) δ: 4.55 (1H, d, J=7.3 Hz), 5.09 (2H, s), 5.14 (2H, m),7.22-7.35 (15H, m), 9.62 (1H, s).

Second Step

Compound 175A (1.30 g, 4.59 mmol), 3-aminopropan-1-ol (379 mg, 5.05mmol) and sodium sulfate (3.26 g, 22.4 mmol) were added to toluene (40ml), and the mixture was stirred at room temperature for 1 hour. Boc₂O(1.17 ml, 5.05 mmol) was added at room temperature, and the mixture wasstirred as it was for 18 hours. Boc₂O (1.17 ml, 5.05 mmol) and sodiumsulfate (3.26 g, 22.4 mmol) were added, and the mixture was stirred for60 hours. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure. The resulting crude product waspurified by silica gel column chromatography (n-hexane-ethyl acetate,1:1, v/v) to obtain 635 mg of 175B as a colorless solid.

Third Step

Compound 175B (632 mg, 1.22 mmol) and palladium-active carbon (10%, wet,100 mg) were added to ethanol (10 ml) and THF (5 ml), and the mixturewas stirred at room temperature for 3 hours. After filtration withCelite, the solvent was concentrated under reduced pressure to obtain502 mg of 175C as a colorless oily substance.

¹H-NMR (CDCl₃) δ: 1.45 (9H, s), 1.77 (2H, m), 3.18-3.27 (1H, m),3.43-3.51 (1H, m), 4.04 (4H, m), 4.92 (1H, d, J=4.7 Hz), 7.28 (10H, m).

Fourth Step

Dimethyl 3-(benzyloxy)-4-oxo-4H-pyran-2,5-dicarboxylate (390 mg, 1.22mmol) and 175C (468 mg, 1.22 mmol) were added to toluene (5 ml), and themixture was stirred at 100° C. for 2 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (n-hexane-ethyl acetate,1:1, v/v) to obtain 391 mg of Compound 175D as a pale yellow gummysubstance.

Fifth Step

To Compound 175D (388 mg, 0.568 mmol) was added 4N HCl (ethyl acetatesolution, 4 ml). After stirring at room temperature for 1 hour, thesolvent was distilled off under reduced pressure. Subsequently, toluene(4 ml) and 3-aminopropan-1-ol (0.0870 ml, 1.14 mmol) were added, and themixture was stirred at 80° C. for 5 hours. After the solvent wasdistilled off under reduced pressure, the resulting crude product waspurified by silica gel column chromatography (chloroform-methanol, 98:2,v/v) to obtain 57 mg of Compound 175E as a yellow gummy substance and 44mg of Compound 175F as a brown gummy substance.

175E: ¹H-NMR (CDCl₃) δ: 1.91-2.00 (2H, m), 2.87 (1H, m), 3.78 (3H, s),3.87-4.15 (3H, m), 4.61 (1H, d, J=12.1 Hz), 4.78 (2H, m), 5.33 (1H, d,J=10.2 Hz), 5.63 (1H, d, J=10.2 Hz), 6.95 (2H, m), 7.13-7.53 (12H, m),7.76 (2H, m)

175F: ¹H-NMR (CDCl₃) δ: 1.83-1.97 (2H, m), 3.12-3.22 (1H, m), 3.50 (1H,m), 3.85 (3H, s), 3.90 (1H, m), 4.34-4.40 (1H, m), 4.74 (1H, d, J=8.6Hz), 4.84-4.89 (1H, m), 5.09 (1H, d, J=3.3 Hz), 5.15 (1H, d, J=9.9 Hz),5.26 (1H, d, J=9.6 Hz), 7.08-7.50 (13H, m), 7.65-7.77 (3H, m).

Sixth Step

Compound 175E (57 mg, 0.10 mmol) was dissolved in THF (0.5 ml) andethanol (0.5 ml), a 2N aqueous sodium hydroxide solution (0.25 ml, 0.50mmol) was added at room temperature, and the mixture was stirred as itwas for 1 hour. To the mixture was added 1N hydrochloric acid, and thiswas extracted with chloroform, and dried with sodium sulfate. Thesolvent was distilled off under reduced pressure, and the resultingcrude product was purified by silica gel column chromatography(chloroform-methanol, 98:2, v/v) to obtain Compound 175G.

Seventh Step

Trifluoroacetic acid (1 ml) was added to Compound 175G obtained in Sixthstep, and the mixture was stirred at room temperature for 1 hour. Afterconcentration under reduced pressure, a pH was adjusted to 3 with anaqueous sodium bicarbonate solution and 2N hydrochloric acid, and thiswas extracted with chloroform, and dried with sodium sulfate. After thesolvent was distilled off under reduced pressure,chloroform-methanol-ethyl ether were added, and the precipitated solidwas filtered off to obtain 11 mg of Compound 175 as a colorless solid.

¹H-NMR (DMSO-d6) δ: 1.50 (1H, d, J=13.1 Hz), 1.79 (1H, m), 3.17 (1H, m),3.86 (1H, t, J=11.0 Hz), 4.03 (1H, dd, J=10.8, 4.1 Hz), 4.46 (1H, d,J=12.0 Hz), 4.53 (1H, dd, J=12.7, 4.2 Hz), 4.84 (1H, s), 5.85 (1H, d,J=11.7 Hz), 7.22 (7H, m), 7.44 (2H, t, J=7.6 Hz), 7.65 (2H, d, J=7.3Hz), 8.14 (1H, s), 12.75 (1H, s), 15.33 (1H, brs).

MS: m/z=447 [M+H]⁺.

Example 28 Measurement of Powder X-Ray Diffraction Pattern

The powder X-ray diffraction measurement of the crystal obtained in eachExample was performed under the following measurement conditionaccording to a method of powder X-ray diffraction measurement describedin a general test method of Japanese Pharmacopoeia.

(Apparatus)

D-8Discover manufactured by Bruker

(Operation Method)

A sample was measured under the following conditions.

Measurement method: reflection methodKind of light source: Cu tubeWavelength used: CuKα-rayTube current: 40 mATube voltage: 40 KvSample plate: glassX-ray incident angle: 3° and 12°

Test Example 1 Measurement of Cap-Dependant Endonuclease (CEN)Inhibitory Activity 1) Preparation of Substrate

30merRNA(5′-pp-[m2′-O]GAA UAU(-Cy3) GCA UCA CUA GUA AGC UUU GCUCUA-BHQ2-3′: manufactured by Japan Bioservice) in which G at a 5′ end isdiphosphate-modified, a hydroxy group at 2′ position ismethoxylation-modified, U sixth from a 5′ end is labelled with Cy3, anda 3′ end is labelled with BHQ2 was purchased, and a cap structure wasadded using ScriptCap system manufactured by EPICENTRE (a product wasm7G [5′]-ppp-[5′] [m2′-O]GAA UAU(-Cy3) GCA UCA CUA GUA AGC UUU GCUCUA(-BHQ2)-3′). This was separated and purified by denaturedpolyacrylamide gel electrophoresis, and used as a substrate.

2) Preparation of Enzyme

RNP was prepared from a virus particle using standard method (ReferenceDocument: VIROLOGY (1976) 73, p 327-338 OLGA M. ROCHOVANSKY).Specifically, A/WSN/33 virus (1×10³ PFU/mL, 200 μL) was innoculated in a10 days old embryonated chicken egg. After incubation at 37° C. for 2days, the allantoic fluid of the chicken egg was recovered. A virusparticle was purified by ultracentrifugation using 20% sucrose,solubilized using TritonX-100 and lysolecithin, and an RNP fraction(50-70% glycerol fraction) was collected by ultracentrifugation using a30-70% glycerol density gradient, and was used as an enzyme solution(containing approximately 1 nM PB1•PB2•PA complex).

3) Enzymatic Reaction

An enzymatic reaction solution (2.54) (composition: 53 mMTris-hydrochloride (pH 7.8), 1 mM MgCl₂, 1.25 mM dithiothreitol, 80 mMNaCl, 12.5% glycerol, enzyme solution 0.15 μL) was dispensed into a384-well plate made of polypropylene. Then, 0.5 μL of a test substancesolution which had been serially diluted with dimethyl sulfoxide (DMSO)was added to the plate. As a positive control (PC) or a negative control(NC), 0.5 μL. of DMSO was added to the plate respectively. Each platewas mixed well. Then, 2 μL of a substrate solution (1.4 nM substrateRNA, 0.05% Tween20) was added to initiate a reaction. After roomtemperature incubation for 60 minutes, 1 μL of the reaction solution wascollected and added to 10 μL of a Hi-Di formamide solution (containingGeneScan 120 Liz Size Standard as a sizing marker: manufactured byApplied Biosystem (ABI)) in order to stop the reaction. For NC, thereaction was stopped in advance by adding EDTA (4.5 mM) beforeinitiation of the reaction (all concentrations described above are finalconcentrations).

3) Measurement of Inhibition Ratio (IC₅₀ Value)

The solution for which the reaction was stopped was heated at 85° C. for5 minutes, rapidly cooled on ice for 2 minutes, and analyzed with an ABIPRIZM 3730 genetic analyzer. A peak of the cap-dependent endonucleaseproduct was quantitated by analysis software ABI Genemapper, a CENreaction inhibition ratio (%) of a test compound was obtained by settingfluorescent intensities of PC and NC to be 0% inhibition and 100%inhibition, respectively, an IC₅₀ value was obtained using curve fittingsoftware (XLfit2.0:Model 205 (manufactured IDBS etc.)). The IC₅₀ valuesof test substances are shown in Table 1.

Test Example 2 CPE Inhibitory Effect Confirming Assay <Material>

2% FCS E-MEM (prepared by adding kanamycin and FCS to MEM (MinimumEssential Medium) (Invitrogen))0.5% BSA E-MEM (prepared by adding kanamycin and BSA to MEM (MinimumEssential Medium) (Invitrogen))

HBSS (Hanks' Balanced Salt Solution)

MDBK cell

Cells were adjusted to the appropriate cell number (3×10⁵/mL) with 2%FCS E-MEM.

MDCK cell

After washing with HBSS two times, cells were adjusted to theappropriate cell number (5×10⁵/mL) with 0.5% BSA E-MEM.

Trypsin Solution

Trypsin from porcine pancreas (SIGMA) was dissolved in PBS(−), andfiltrated with a 0.45 μm filter.

EnVision (Perkin Elmer) WST-8 Kit (Kishida Chemical Co., Ltd.)

10% SDS solution

<Operation Procedure> Dilution and Dispensation of Test Sample

As a culture medium, 2% FCS E-MEM was used at the use of MDBK cells, and0.5% BSA E-MEM was used at the use of MDCK cells. Hereinafter, fordiluting virus, cells and a test sample, the same culture medium wasused.

A test sample was diluted with a culture medium to a appropriateconcentration in advance, and then 2 to 5-fold serial dilution on a 96well plate (50 μL/well) was prepared. Two plate, one for measuringanti-Flu activity and the other for measuring cytotoxity, were prepared.Each assay was performed triplicate for each drug.

At the use of MDCK cells, trypsin was added to the cells to be a finalconcentration of 3 μg/mL only for measuring anti-Flu activity.

Dilution and Dispensation of Influenza Virus

An influenza virus was diluted with a culture medium to a appropriateconcentration in advance, and each 50 μL/well was dispensed on a 96-wellplate containing a test substance. Each 50 μL/well of a culture mediumwas dispensed on a plate containing a test substance for measuringcytotoxity.

Dilution and Dispensation of Cell

Each 100 μL/well of cells which had been adjusted to the appropriatecell number was dispensed on a 96 well plate containing a testsubstance. This was mixed with a plate mixer, and incubated in a CO₂incubator for 3 days for measuring anti-Flu activity and measuringcytotoxity.

Dispensation of WST 8

The cells in 96-well plate which had been incubated for 3 days wasobserved visually under a microscope, and appearance of the cells, thepresence or absence of a crystal of test substance were checked. Thesupernatant was removed so that the cells were not absorbed from theplate.

WST-8 Kit was diluted 10-fold with a culture medium, and each 1000, wasdispensed into each well. After mixing with a plate mixer, cells wereincubated in a CO₂ incubator for 1 to 3 hours.

After incubation, regarding the plate for measuring anti-Flu activity,each 10 μL/well of a 10% SDS solution was dispensed in order toinactivate a virus. Measurement of absorbance

After the 96-well plate was mixed, absorbance was measured with EnVisionat two wavelengths of 450 nm/620 nm.

<Calculation of Each Measurement Item Value>

The value was calculated using Microsoft Excel or a program having theequivalent calculation and processing ability, based on the followingcalculation equation.

Calculation of effective concentration to achieve 50% CPE inhibition(EC50)

EC50=10̂Z

Z=(50%−High %)/(High %−Low %)×{log(High conc.)−log(Low conc.)}+log(Highconc.)

IC₅₀ values of test substances are shown in Table 1.

TABLE 1 CEN IC₅₀ CPE IC₅₀ Examle No. Compound No. (μM) (μM) 14 43 0.0781.41 21 128 0.063 0.416 27 175 0.132 0.102

The test substances exhibited the high cap-dependent endonuclease (CEN)inhibitory activity, and exhibited the high CPE inhibitory effect. Thesesubstances can be medicaments useful as a therapeutic agent and/orprophylactic agent of a symptom and/or a disease induced by infectionwith influenza virus.

Therefore, it can be said that the substance and production method ofthe present invention are an intermediate substance and a productionmethod useful for efficiently producing substances which can be used asa medicament.

1-18. (canceled)
 19. A crystal of a compound shown by formula (1D) or asolovate thereof:

wherein Me is a methyl group, and Bn is a benzyl group, wherein thecompound has a peak at a diffraction angle (28): 7.9°±0.2°, 10.0°±0.2°,11.5°±0.2°, 20.0°±0.2°, 23.4°±0.2° and 34.0°±0.2° in a powder X-raydiffraction spectrum.
 20. A crystal of a compound represented by formula(2D) or a solvate thereof:

wherein Me is a methyl group, Et is an ethyl group, and Bn is a benzylgroup, wherein the compound has a peak at a diffraction angle (28):17.6°±0.2°, 25.2°±0.2°, 26.4°±0.2° and 28.1°±0.2° in a powder X-raydiffraction spectrum.
 21. A crystal of a compound shown by formula (9C′)or a solvate thereof:

wherein Me is a methyl group, and Bn is a benzyl group, wherein thecompound has a peak at a diffraction angle (28): 14.2°±0.2°, 16.0°±0.2°,22.0°±0.2°, 22.2°±0.2°, 24.4°±0.2° and 25.9°±0.2° in a powder X-raydiffraction spectrum.
 22. A crystal according to claim 19, which ischaracterized by a powder X-ray diffraction spectrum which issubstantially consistent with FIG.
 1. 23. A crystal according to claim20, which is characterized by a powder X-ray diffraction spectrum whichis substantially consistent with FIG.
 2. 24. A crystal according toclaim 21, which is characterized by a powder X-ray diffraction spectrumwhich is substantially consistent with FIG. 3.